NIH References of Lab rec.

Goat anti Adenovirus
MBS315064-1mL MyBiosource 1mL 885 EUR
Goat anti Adenovirus
MBS315064-5x1mL MyBiosource 5x1mL 3815 EUR
Goat anti Adenovirus
MBS315065-1mL MyBiosource 1mL 770 EUR
Goat anti Adenovirus
MBS315065-5x1mL MyBiosource 5x1mL 3290 EUR
MOUSE ANTI ADENOVIRUS
MBS213040-1mg MyBiosource 1mg 805 EUR
MOUSE ANTI ADENOVIRUS
MBS213040-5x1mg MyBiosource 5x1mg 3440 EUR
Mouse Adenovirus-1 FL Antigen, Adenovirus
MBS412249-5xHalfELISAPlatesEquivalent5x48Reactions MyBiosource 5xHalfELISAPlatesEquivalent(5x48Reactions) 1500 EUR
Mouse Adenovirus-2 K87 Antigen, Adenovirus
MBS412250-5xHalfELISAPlatesEquivalent5x48Reactions MyBiosource 5xHalfELISAPlatesEquivalent(5x48Reactions) 1500 EUR
MOUSE ANTI ADENOVIRUS HEXON
MBS211445-02mg MyBiosource 0.2mg 515 EUR
MOUSE ANTI ADENOVIRUS HEXON
MBS211445-5x02mg MyBiosource 5x0.2mg 2145 EUR
MOUSE ANTI ADENOVIRUS HEXON
MBS211446-02mg MyBiosource 0.2mg 515 EUR
MOUSE ANTI ADENOVIRUS HEXON
MBS211446-5x02mg MyBiosource 5x0.2mg 2145 EUR
MOUSE ANTI ADENOVIRUS HEXON
MBS211447-02mg MyBiosource 0.2mg 515 EUR
MOUSE ANTI ADENOVIRUS HEXON
MBS211447-5x02mg MyBiosource 5x0.2mg 2145 EUR
MOUSE ANTI ADENOVIRUS HEXON
MBS211443-02mg MyBiosource 0.2mg 515 EUR
MOUSE ANTI ADENOVIRUS HEXON
MBS211443-5x02mg MyBiosource 5x0.2mg 2145 EUR
MOUSE ANTI ADENOVIRUS HEXON:FITC
MBS211132-01mg MyBiosource 0.1mg 640 EUR
MOUSE ANTI ADENOVIRUS HEXON:FITC
MBS211132-5x01mg MyBiosource 5x0.1mg 2700 EUR
anti-Coxsackie Adenovirus Receptor
YF-PA11220 Abfrontier 50 ug 435.6 EUR
anti-Coxsackie Adenovirus Receptor
YF-PA11221 Abfrontier 100 ug 483.6 EUR
ADENOVIRUS
GWB-20CCE1 GenWay Biotech 1 ml Ask for price
Adenovirus
GWB-EE7DB9 GenWay Biotech 1 ml Ask for price
Adenovirus
GWB-C61E23 GenWay Biotech 0.1 mg Ask for price
Adenovirus
PCR-H501-PCRH50148D Bioingentech PCR-H501-48D 230 EUR
Adenovirus
PCR-H501-PCRH50196D Bioingentech PCR-H501-96D 312 EUR
Adenovirus
Oneq-H501-OneqH501100D Bioingentech Oneq-H501-100D 515 EUR
Adenovirus
Oneq-H501-OneqH501150D Bioingentech Oneq-H501-150D 594 EUR
Adenovirus
Oneq-H501-OneqH50150D Bioingentech Oneq-H501-50D 413 EUR
Adenovirus
1411 Virostat each 270 EUR
Adenovirus
MBS655204-05mg MyBiosource 0.5mg 1850 EUR
Adenovirus
MBS655204-5x05mg MyBiosource 5x0.5mg 8175 EUR
Adenovirus
MBS6506950-01mg MyBiosource 0.1mg 580 EUR
Adenovirus
MBS6506950-5x01mg MyBiosource 5x0.1mg 2470 EUR
Adenovirus
MBS6506952-01mg MyBiosource 0.1mg 575 EUR
Adenovirus
MBS6506952-5x01mg MyBiosource 5x0.1mg 2440 EUR
Adenovirus
MBS6506953-01mg MyBiosource 0.1mg 605 EUR
Adenovirus
MBS6506953-5x01mg MyBiosource 5x0.1mg 2575 EUR
Adenovirus
MBS320083-01mg MyBiosource 0.1mg 355 EUR
Adenovirus
MBS320083-5x01mg MyBiosource 5x0.1mg 1500 EUR
Adenovirus
MBS320084-01mg MyBiosource 0.1mg 355 EUR
Adenovirus
MBS320084-5x01mg MyBiosource 5x0.1mg 1500 EUR
Adenovirus
MBS320085-01mg MyBiosource 0.1mg 355 EUR
Adenovirus
MBS320085-5x01mg MyBiosource 5x0.1mg 1500 EUR
Adenovirus
MBS320086-01mg MyBiosource 0.1mg 395 EUR
Adenovirus
MBS320086-5x01mg MyBiosource 5x0.1mg 1685 EUR
Adenovirus
MBS320087-01mg MyBiosource 0.1mg 355 EUR
Adenovirus
MBS320087-5x01mg MyBiosource 5x0.1mg 1500 EUR
Adenovirus
MBS320088-01mg MyBiosource 0.1mg 395 EUR
Adenovirus
MBS320088-5x01mg MyBiosource 5x0.1mg 1685 EUR
Adenovirus
MBS320089-01mg MyBiosource 0.1mg 355 EUR
Adenovirus
MBS320089-5x01mg MyBiosource 5x0.1mg 1500 EUR
Adenovirus
MBS320090-01mg MyBiosource 0.1mg 355 EUR
Adenovirus
MBS320090-5x01mg MyBiosource 5x0.1mg 1500 EUR
Adenovirus
MBS320091-01mg MyBiosource 0.1mg 355 EUR
Adenovirus
MBS320091-5x01mg MyBiosource 5x0.1mg 1500 EUR
Adenovirus
MBS370003-01mLConcentrate MyBiosource 0.1mL(Concentrate) 240 EUR
Adenovirus
MBS370003-05mLConcentrate MyBiosource 0.5mL(Concentrate) 465 EUR
Adenovirus
MBS370003-15mLRTU MyBiosource 15mL(RTU) 775 EUR
Adenovirus
MBS370003-3mLRTU MyBiosource 3mL(RTU) 300 EUR
Adenovirus
MBS370003-7mLRTU MyBiosource 7mL(RTU) 465 EUR
Adenovirus
MBS324088-1mL MyBiosource 1mL 315 EUR
Adenovirus
MBS324088-5x1mL MyBiosource 5x1mL 1320 EUR
Adenovirus
MBS324089-1mL MyBiosource 1mL 325 EUR
Adenovirus
MBS324089-5x1mL MyBiosource 5x1mL 1375 EUR
Adenovirus
MBS324090-1mL MyBiosource 1mL 345 EUR
Adenovirus
MBS324090-5x1mL MyBiosource 5x1mL 1465 EUR

Compare ELISA lab reagents for research





Suppliers for Lab recombinants

pRK5 Plasmid

PVT48959 Nova Lifetech 2ug

ePop Plasmid

PVT50044 Nova Lifetech 2ug

dsDNA plasmid

30R-AD006 Fitzgerald 1 mg
Description: Double stranded DNA plasmid

dsDNA plasmid

MBS537931-1mg MyBiosource 1mg

dsDNA plasmid

MBS537931-5x1mg MyBiosource 5x1mg

TRPM4 Plasmid

PVT51226 Nova Lifetech 2ug

pLysS Plasmid

PVT5702 Nova Lifetech 2ug

Rat Cholesterol ELISA ELISA

E01A11128 BlueGene 96T 700 EUR

Goat Cholesterol ELISA ELISA

E01A46041 BlueGene 96T 700 EUR

Mouse Cholesterol ELISA ELISA

E01A19869 BlueGene 96T 700 EUR

Human Cholesterol ELISA ELISA

E01A2368 BlueGene 96T 700 EUR

Sheep Cholesterol ELISA ELISA

E01A98335 BlueGene 96T 700 EUR

Monkey Cholesterol ELISA ELISA

E01A72187 BlueGene 96T 700 EUR

Canine Cholesterol ELISA ELISA

E01A63475 BlueGene 96T 700 EUR

Rabbit Cholesterol ELISA ELISA

E01A28609 BlueGene 96T 700 EUR

Our used TESTs in Pubmed.

NCC Antibody, Anti-NCC Antibody

MBS805073-01mg MyBiosource 0.1mg 525 EUR

NCC Antibody, Anti-NCC Antibody

MBS805073-5x01mg MyBiosource 5x0.1mg 1830 EUR

Tau (Ab-262) Antibody Antibody

E11-7239B EnoGene 100μg/100μl 225 EUR

Antibody Pair to APOA5 Antibody

E10-20042 EnoGene 100μg/100μl 225 EUR

CLCN5 Antibody / CIC-5 antibody

RQ6462 NSJ Bioreagents 100ug 356.15 EUR

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Suppliers for Lab Assays

HIV-2 gp36

hiv-104 ProSpec Tany 100µg
Description: Synthetic HIV-2 gp36

HIV-1 gp41

hiv-112 ProSpec Tany 100µg
Description: Recombinant HIV-1 gp41

HIV-2 gp32

hiv-134 ProSpec Tany 100µg
Description: Recombinant HIV-2 gp32

HIV-2 gp36

MBS8506649-01mg MyBiosource 0.1mg

HIV-2 gp36

MBS8506649-5x01mg MyBiosource 5x0.1mg

HIV-1 gp120

E62H004 EnoGene 100ug

HIV-1 gp160

E62H00401 EnoGene 100ug

Monoclonal GR monoclonal antibody

AMM00029G Leading Biology 0.05mg 633.6 EUR

Monoclonal TBP monoclonal antibody

APR13720G Leading Biology 0.1ml 633.6 EUR

Monoclonal EZH2 monoclonal antibody

AMM00030G Leading Biology 0.05mg 633.6 EUR

Monoclonal Rsf1 monoclonal antibody

AMM07673G Leading Biology 0.05mg 633.6 EUR

Monoclonal Rsf1 monoclonal antibody

AMM07674G Leading Biology 0.1ml 633.6 EUR

Monoclonal HDAC2 monoclonal antibody

AMM00031G Leading Biology 0.05mg 633.6 EUR

Monoclonal SirT1 monoclonal antibody

APR09951G Leading Biology 0.05mg 580.8 EUR

Monoclonal SirT1 monoclonal antibody

APR09952G Leading Biology 0.1ml 580.8 EUR

Our used polyclonals in Pubmed.

Anti-Anti-SEPT7 Antibody antibody

STJ116214 St John's Laboratory 100 µl 332.4 EUR

Anti-Anti-SEPT4 Antibody antibody

STJ112276 St John's Laboratory 100 µl 332.4 EUR

Anti-Anti-SEPT8 Antibody antibody

STJ117206 St John's Laboratory 100 µl 332.4 EUR

Anti-Anti-SEPT2 Antibody antibody

STJ28365 St John's Laboratory 100 µl 332.4 EUR

Anti-Anti-SEPT3 Antibody antibody

STJ118990 St John's Laboratory 100 µl 332.4 EUR

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Suppliers for Lab ELISAs

Equine Eye cDNA

ED-106 Zyagen 30 reactions

Cat Tongue cDNA

FD-105 Zyagen 30 Reactions

Cat Rectum cDNA

FD-312 Zyagen 30 Reactions

Cat Testis cDNA

FD-401 Zyagen 30 Reactions

Cat Uterus cDNA

FD-411 Zyagen 30 Reactions

Cat Spleen cDNA

FD-701 Zyagen 30 Reactions

Cat Thymus cDNA

FD-702 Zyagen 30 Reactions

REC-1

ABC-TC0956 AcceGen 1 vial Ask for price

REC-2615 (HCl)

530142 MedKoo Biosciences 10.0mg 295 EUR

Rec FLA-ST

tlrl-flic-10 InvivoGen FR 10 µg 295.05 EUR

Rec FLA-ST

tlrl-flic-50 InvivoGen FR 50 µg 731.85 EUR

rec EGF (human)

4030572.01 Bachem 0.1 mg 102.27 EUR

rec EGF (human)

4030572.05 Bachem 0.5 mg 271.85 EUR

rec EGF (human)

H-7490.0100 Bachem 0.1mg 194.4 EUR

rec EGF (human)

H-7490.0500 Bachem 0.5mg 457.2 EUR

Our used monoclonals in Pubmed.

CNX ELISA Kit| Rat Calnexin ELISA Kit

EF017638 Lifescience Market 96 Tests 826.8 EUR

GHRL ELISA Kit| Rat Ghrelin ELISA Kit

EF017639 Lifescience Market 96 Tests 826.8 EUR

MT ELISA Kit| Rat Melatonin ELISA Kit

EF017898 Lifescience Market 96 Tests 826.8 EUR

NPHN ELISA Kit| Rat Nephrin ELISA Kit

EF017918 Lifescience Market 96 Tests 826.8 EUR

E2 ELISA Kit| Rat Estradiol ELISA Kit

EF018157 Lifescience Market 96 Tests 826.8 EUR

Compare Appoptosis lab reagents for research






Suppliers for Lab monoclonals

Rabbit pAbPC3 Rabbit pAb

A18370-100ul Abclonal 100 ul

Rabbit pAbPC3 Rabbit pAb

A18370-200ul Abclonal 200 ul

Rabbit pAbPC3 Rabbit pAb

A18370-20ul Abclonal 20 ul

Rabbit pAbPC3 Rabbit pAb

A18370-50ul Abclonal 50 ul

Rabbit pAbPC4 Rabbit pAb

A5948-100ul Abclonal 100 ul

Rabbit pAbPC4 Rabbit pAb

A5948-200ul Abclonal 200 ul

Rabbit pAbPC4 Rabbit pAb

A5948-20ul Abclonal 20 ul

H2B Antibody Antibody

E11-184659 EnoGene 100ug/100ul 225 EUR

Lck antibody Antibody

GWB-250026 GenWay Biotech 0.05 ml Ask for price

anti- Antibody^Polyclonal antibody control antibody

LSMab09882 Lifescience Market 100 ug 525.6 EUR

CD11b Antibody Antibody

E19-2911-1 EnoGene 50ug/50ul 145 EUR

CD11b Antibody Antibody

E19-2911-2 EnoGene 100ug/100ul 225 EUR

ZNF98 Antibody Antibody

E36403 EnoGene 100μg 275 EUR

HSP60 Antibody Antibody

E8M1007-4 EnoGene 100ul 225 EUR

ASAP1 antibody Antibody

DF8746 Affbiotech 200ul 420 EUR

Our used polyclonals in Pubmed.

rec PDGF AA (human)

H-7640.0002 Bachem 2.0µg 194.4 EUR

rec PDGF AA (human)

H-7640.0010 Bachem 10.0µg 457.2 EUR

rec IGF-I (human)

H-5555.0050 Bachem 50.0µg 290.4 EUR

rec IGF-I (human)

H-5555.0100 Bachem 100.0µg 457.2 EUR

rec IGF-I (human)

H-5555.0500 Bachem 0.5mg 1720.8 EUR

antibody Lab Reagents for Research






Promoted Lab rec.

H2B Antibody Antibody

E11-184659 EnoGene 100ug/100ul 225 EUR

Lck antibody Antibody

GWB-250026 GenWay Biotech 0.05 ml Ask for price

anti- Antibody^Polyclonal antibody control antibody

LSMab09882 Lifescience Market 100 ug 525.6 EUR

CD11b Antibody Antibody

E19-2911-1 EnoGene 50ug/50ul 145 EUR

CD11b Antibody Antibody

E19-2911-2 EnoGene 100ug/100ul 225 EUR

ASAP1 antibody Antibody

DF8746 Affbiotech 200ul 420 EUR

REC-1

ABC-TC0956 AcceGen 1 vial Ask for price

REC-2615 (HCl)

530142 MedKoo Biosciences 10.0mg 295 EUR

Rec FLA-ST

tlrl-flic-10 InvivoGen FR 10 µg 295.05 EUR

Rec FLA-ST

tlrl-flic-50 InvivoGen FR 50 µg 731.85 EUR

rec EGF (human)

4030572.01 Bachem 0.1 mg 102.27 EUR

rec EGF (human)

4030572.05 Bachem 0.5 mg 271.85 EUR

rec EGF (human)

H-7490.0100 Bachem 0.1mg 194.4 EUR

Our used recombinants in Pubmed.

TAGLN Recombinant Protein (Human) (Recombinant- Tag)

RP030886 ABM 100 ug Ask for price

TAGAP Recombinant Protein (Human) (Recombinant- Tag)

RP043930 ABM 100 ug Ask for price

CTAGE5 Recombinant Protein (Mouse) (Recombinant- Tag)

RP126449 ABM 100 ug Ask for price

CTAGE5 Recombinant Protein (Mouse) (Recombinant- Tag)

RP126452 ABM 100 ug Ask for price

CTAGE5 Recombinant Protein (Mouse) (Recombinant- Tag)

RP126455 ABM 100 ug Ask for price

CTAGE1 Recombinant Protein (Human) (Recombinant- Tag)

RP008266 ABM 100 ug Ask for price

CTAGE5 Recombinant Protein (Human) (Recombinant- Tag)

RP008269 ABM 100 ug Ask for price

CTAGEP Recombinant Protein (Human) (Recombinant- Tag)

RP008275 ABM 100 ug Ask for price

Tagap1 Recombinant Protein (Mouse) (Recombinant Tag)

RP177158 ABM 100 ug Ask for price

TAGLN2 Recombinant Protein (Mouse) (Recombinant Tag)

RP177164 ABM 100 ug Ask for price

A CRISPR toolbox for generating intersectional genetic mouse models for functional, molecular, and anatomical circuit mapping

In addition to single recombinase systems, the expression of two recombinases in distinct, but partially overlapping, populations allows for more defined target expression. Although the application of this method is becoming increasingly popular, its experimental implementation has been broadly restricted to manipulations of a limited set of common alleles that are often commercially produced at great expense, with costs and technical challenges associated with the production of intersectional mouse lines hindering customized approaches to many researchers Joblinks Human ADAM10 cDNA. Here, we present a simplified CRISPR toolkit for rapid, inexpensive, and facile intersectional allele production.
Results: Briefly, we produced 7 intersectional mouse lines using a dual recombinase system, one mouse line with a single recombinase system, and three embryonic stems (ES) cell lines that are designed to study the way functional, molecular, and anatomical features relate to each other in building circuits that underlie physiology and behavior.
As a proof-of-principle, we applied three of these lines to different neuronal populations for anatomical mapping and functional in vivo investigation of respiratory control.
We also generated a mouse line with a single recombinase-responsive allele that controls the expression of the calcium sensor Twitch-2B. This mouse line was applied globally to study the effects of follicle-stimulating hormone (FSH) and luteinizing hormone (LH) on calcium release in the ovarian follicle.
Conclusions: The lines presented here are representative examples of outcomes possible with the successful application of our genetic toolkit for the facile development of diverse, modifiable animal models. This toolkit will allow labs to create single or dual recombinase effector lines easily for any cell population or subpopulation of interest when paired with the appropriate Cre and FLP recombinase mouse lines or viral vectors. We have made our tools and derivative intersectional mouse and ES cell lines openly available for non-commercial use through publicly curated repositories for plasmid DNA, ES cells, and transgenic mouse lines.

An enhanced method for nucleic acid detection with CRISPR-Cas12a using phosphorothioate modified primers and optimized gold-nanopaticle strip

CRISPR-Cas12a system has been shown promising for nucleic acid diagnostics due to its rapid, portable and accurate features. However, cleavage of the amplicons and primers by the cis– and trans-activity of Cas12a hinders the attempts to integrate the amplification and detection into a single reaction. Through phosphorothioate modification of primers, we realized onepot detection with high sensitivity using plasmids of SARS-CoV-2, HPV16 and HPV18. We also identified the activated Cas12a has a much higher affinity to C nucleotide-rich reporter than others.
By applying such reporters, the reaction time required for a lateral-flow readout was significantly reduced. Furthermore, to improve the specificity of the strip-based assay, we created a novel reporter and, when combined with a customized gold-nanopaticle strip, the readout was greatly enhanced owing to the elimination of the nonspecific signal.
This established system, termed Targeting DNA by Cas12a-based Eye Sight Testing in an One-pot Reaction (TESTOR), was validated using clinical cervical scrape samples for human papillomaviruses (HPVs) detection. Our system represents a general approach to integrating nucleic acid amplification and detection into a single reaction in CRISPR-Cas systems, highlighting its potential as a rapid, portable and accurate detection platform of nucleic acids.

Detection of plasmid contigs in draft genome assemblies using customized Kraken databases

Plasmids play an important role in bacterial evolution and mediate horizontal transfer of genes including virulence and antimicrobial resistance genes. Although short-read sequencing technologies have enabled large-scale bacterial genomics, the resulting draft genome assemblies are often fragmented into hundreds of discrete contigs. Several tools and approaches have been developed to identify plasmid sequences in such assemblies, but require trade-off between sensitivity and specificity. Here we propose using the Kraken classifier, together with a custom Kraken database comprising known chromosomal and plasmid sequences of Klebsiella pneumoniae species complex (KpSC), to identify plasmid-derived contigs in draft assemblies.
We assessed performance using Illumina-based draft genome assemblies for 82 KpSC isolates, for which complete genomes were available to supply ground truth. When benchmarked against five other classifiers (Centrifuge, RFPlasmid, mlplasmids, PlaScope and Platon), Kraken showed balanced performance in terms of overall sensitivity and specificity (90.8 and 99.4 %, respectively, for contig count; 96.5 and >99.9 %, respectively, for cumulative contig length), and the highest accuracy (96.8% vs 91.8-96.6% for contig count; 99.8% vs 99.0-99.7 % for cumulative contig length), and F1-score (94.5 % vs 84.5-94.1 %, for contig count; 98.0 % vs 88.9-96.7 % for cumulative contig length). Kraken also achieved consistent performance across our genome collection. Furthermore, we demonstrate that expanding the Kraken database with additional known chromosomal and plasmid sequences can further improve classification performance. Although we have focused here on the KpSC, this methodology could easily be applied to other species with a sufficient number of completed genomes.

Exploiting heterologous and endogenous CRISPR-Cas systems for genome editing in the probiotic Clostridium butyricum

Clostridium butyricum has been widely used as a probiotic for humans and food animals. However, the mechanisms of beneficial effects of C. butyricum on the host remain poorly understood, largely due to the lack of high-throughput genome engineering tools. Here, we report the exploitation of heterologous Type II CRISPR-Cas9 system and endogenous Type I-B CRISPR-Cas system in probiotic C. butyricum for seamless genome engineering. Although successful genome editing was achieved in C. butyricum when CRISPR-Cas9 system was employed, the expression of toxic cas9 gene result in really poor transformation, spurring us to develop an easy-applicable and high-efficient genome editing tool.
Therefore, the endogenous Type I-B CRISPR-Cas machinery located on the megaplasmid of C. butyricum was co-opted for genome editing. In vivo plasmid, interference assays identified that ACA and TAA were functional protospacer adjacent motif (PAM) sequences needed for site-specific CRISPR attacking. Using the customized endogenous CRISPR-Cas system, we successfully deleted spo0A and aldh genes in C. butyricum, yielding an efficiency of up to 100%.
Moreover, the conjugation efficiency of endogenous CRISPR-Cas system was dramatically enhanced due to the precluding expression of cas9. Altogether, the two approaches developed herein remarkably expand the existing genetic toolbox available for investigation of C. butyricum. This article is protected by copyright. All rights reserved.
ADAM10 (Human)
GT15078 100 ug 631.2 EUR
Human ADAM10 ELISA
KT-5207 96 tests 952 EUR
Human ADAM10 Protein
20-abx262305
  • Ask for price
  • Ask for price
  • Ask for price
  • 10 ug
  • 1 mg
  • 2 µg
Human ADAM10 Protein
abx262305-10mg 10 mg 325 EUR
Human ADAM10 Protein
abx262305-25mg 25 mg 6575 EUR
Human ADAM10 Protein
abx262305-5mg 5 mg 225 EUR
ADAM10 (Human), CF
PR15036CF 20 ug 631.2 EUR
ADAM10 (untagged)-Human ADAM metallopeptidase domain 10 (ADAM10)
SC119437 10 µg Ask for price
ADAM10 siRNA (Human)
MBS8233659-15nmol 15nmol 405 EUR
ADAM10 siRNA (Human)
MBS8233659-30nmol 30nmol 565 EUR

Delivery of superoxide dismutase by TAT and Abalone peptides for the protection of skin cells against oxidative stress

This work aimed to clone, express, purify and evaluate the protective effect antioxidant of this enzyme on skin cells when fused to transactivator of transcription (TAT) protein transduction domain of HIV-1 and Abalone (Ab) peptides to allow cell penetration. TrSOD, TAT-TrSOD-Yfp (fused to yellow fluorescent protein) and Ab-TrSOD were expressed in E.coli and purified as soluble proteins. The cytotoxicity of the enzymes, at the concentrations of 1, 3 and 6 μmol/L, was evaluated for a period of 24 and 48 h of incubation, with no cytotoxic effect on 3T3 fibroblasts. The 3T3 cells were exposed to the oxidant agent tert-butyl hydroperoxide (tBH) and evaluated for ROS generation, in the presence or not of the recombinant enzymes.
TAT-TrSOD-Yfp was able to decrease the generation of ROS in 15% when used in the concentrations of 3 and 6 μmol/L in comparison to the control, but there was no difference in relation to the effect of TrSOD. Ab-TrSOD, when compared to TrSOD, promoted a decrease in the formation of ROS of 19 and 14% at the concentrations of 1 and 6 μmol/L, respectively, indicating that this joplink Recombinant Human Regulator of G-protein form was more effective in reducing oxidative stress compared to SOD without the cell penetrating peptide (CPP).
Together, these results indicate that the fusion of SOD with these CPP increased the antioxidant capacity of fibroblasts, identified by the reduction in the generation of ROS. In addition, such molecules, in the concentrations initially used, were not toxic to the cells, opening perspectives for the development of products for antioxidant protection of the skin that may have therapeutic and cosmetic application. This article is protected by copyright.

Comparative Immunogenicity of the Recombinant Receptor-Binding Domain of Protein S SARS-CoV-2 Obtained in Prokaryotic and Mammalian Expression Systems

The receptor-binding domain (RBD) of the protein S SARS-CoV-2 is considered to be one of the appealing targets for developing a vaccine against COVID-19. The choice of an expression system is essential when developing subunit vaccines, as it ensures the effective synthesis of the correctly folded target protein, and maintains its antigenic and immunogenic properties. Here, we describe the production of a recombinant RBD protein using prokaryotic (pRBD) and mammalian (mRBD) expression systems, and compare the immunogenicity of prokaryotic and mammalian-expressed RBD using a BALB/c mice model.
An analysis of the sera from mice immunized with both variants of the protein revealed that the mRBD expressed in CHO cells provides a significantly stronger humoral immune response compared with the RBD expressed in E.coli cells. A specific antibody titer of sera from mice immunized with mRBD was ten-fold higher than the sera from the mice that received pRBD in ELISA, and about 100-fold higher in a neutralization test. The data obtained suggests that mRBD is capable of inducing neutralizing antibodies against SARS-CoV-2.

Preparation of highly specific monoclonal antibodies against SARS-CoV-2 nucleocapsid protein and the preliminary development of antigen detection test strips

The coronavirus disease 2019 (COVID-19) are outbreaking all over the world. To help fight this disease, it is necessary to establish an effective and rapid detection method. The nucleocapsid (N) protein of Severe Acute Respiratory syndrome Coronavirus 2 (SARS-CoV-2) is involved in viral replication, assembly and immune regulation and plays an important role in the viral life cycle. Moreover, the N protein also could be a diagnostic factor and potential drag target.
Therefore, by synthesizing the N gene sequence of SARS-CoV-2, constructing the pET-28a (+)-N recombinant plasmid, we expressed the N protein in E.coli and obtained 15 mAbs against SARS-CoV-2-N protein by the hybridomas and ascites, then an immunochromatographic test strip method detecting N antigen was established.
In this study, we obtained 14 high-titer and high-specificity monoclonal antibodies, and the test strips exclusively react with the SARS-CoV-2-N protein and no cross-reactivity with other coronavirus and also recognize the recombinant N protein of Delta (B.1.617.2) variant. These mAbs can be used for the early and rapid diagnosis of SARS-CoV-2 infection through serological antigen. This article is protected by copyright. All rights reserved.

Preparation and identification of rat polyclonal antibody against SARS-CoV-2 main protease (Mpro)

Objective To investigate the immunological functions of SARS-CoV-2 main protease (Mpro) in coronavirus disease 2019 (COVID-19), polyclonal antibody against Mpro was developed. Methods A codon-optimized SARS-CoV-2 Mpro gene was synthesized and ligated into a pET-28a vector for construction of a recombinant plasmid named by pET-28a-Mpro. Subsequently, this plasmid was transformed into E.coli Rosetta (DE3) competent cells for Mpro expression in an optimized condition, and then Mpro was purified using a HisTrap chelating column.
The purified Mpro was used as immunogen to inoculate rats and the serum was collected after third immunization cycle. The titer, selectivity and sensitivity of polyclonal antibody against Mpro were analyzed using the ELISA and Western blot analysis. Results An optimized expression condition in E.coli cells for Mpro was determined, and the recombinant Mpro was purified by a HisTrap chelating column. The ELISA and Western blot analysis demonstrated that the highly sensitive polyclonal antibody against Mpro specially recognized the recombinant Mpro, and the titer reached 1:256 000. Conclusion The highly specific polyclonal antibody against SARS-CoV-2 Mpro is successfully prepared, which lays an experimental foundation for investigating the immunological function of Mpro in COVID-19.

Direct enzyme-linked aptamer assay (DELAA) for diagnosis of toxoplasmosis by detection of SAG1 protein in mice and humans

Toxoplasma gondii is a single-celled parasite commonly found in mammals and birds. Diagnosis of toxoplasmosis largely depends on measurements of the antibody and/or antigen and Toxoplasma DNAs due to the presence of tissue dwelling duplicating tachyzoites, or quiescent cysts in latent infection of the parasite. As a major surface antigen of T.gondii tachyzoites, SAG1 is a key molecule for laboratory diagnosis. However, there are no methods available yet for SAG1 detection using aptamer-based technology. Recombinant SAG1 (r-SAG1) of Toxoplasma WH3 strain (type Chinese 1) was expressed in E.coli and subjected to the synthetic oligonucleotide library for selection of nucleic acid aptamers which target the r-SAG1 antigen, with systematic evolution of ligands by exponential enrichment (SELEX) strategy.
The specific aptamers were screened out and used in direct enzyme-linked aptamer assay (DELAA) for detection of native SAG1 (n-SAG1) obtained from tachyzoite lysates, mouse sera of acute infection, and human sera that had been verified for Toxoplasma DNAs by PCR amplification. As results, the soluble r-SAG1 protein was obtained from E.coli lysates by purification and identification with immunoblotting, followed by biotinylation. The selected aptamers were amplified by PCR and DNA sequencing.
The results showed that the aptamer-2, with the highest affinity to n-SAG1 in the sera of animals with minimal difference in the four aptamer candidates, has a high specificity and sensitivity when used in detection of n-SAG1 in the sera of humans when compared with the commercial kit of ELISA for T.gondii circulating antigen test. We concluded that a new direct enzyme-linked aptamer assay (DELAA) was developed for the detection of the n-SAG1 protein of T. gondii. With increased sensitivity and specificity, stability, easy and cheap preparation, the aptamer-based technology is considered an efficient method for the diagnosis of active as well as reactivated toxoplasmosis.
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DETECTION METHODS FOR WESTERN BLOT

Western Blot is a technique commonly used in scientific research laboratories. By means of this technique, the different proteins present in a sample are separated according to their molecular weight by gel electrophoresis, and are subsequently transferred to a membrane to proceed with their identification using specific antibodies.

Although the foundation of the technique remains, over the years new detection methods have been developed in order to obtain more accurate results and also allow a quantitative analysis of proteins.

In this post we bring you a summary of the main detection methods for Western Blot with the advantages and disadvantages of each of them.

DETECTION METHODS FOR WESTERN BLOT

1.- RADIOACTIVE DETECTION METHODS
This type of detection was one of the first methods used to reveal the results of the Western Blot, by labeling the antibodies with radioactive conjugates.

The main advantage of this method lies in its sensitivity, but it has the great drawback that when using radioactive materials there is a risk to the researcher’s health and safety. Furthermore, it is a high-cost technique and its execution is time consuming.

Radioactive detection is not currently among the Western Blot detection methods of choice. In fact, its use is discouraged.

2.- ENZYMATIC DETECTION METHODS
These methods are based on the use of secondary antibodies conjugated to an enzyme that catalyzes a reaction with a specific substrate.

Within this category, detection can be carried out by means of two types of enzymatic reactions:

2.1 COLORIMETRIC DETECTION
In this case, the enzyme bound to the secondary antibody triggers a reaction with the substrate giving rise to a colored precipitate that can be visually identified.

The advantages of this method lie in its speed, simplicity and low economic cost, in addition to not requiring any special equipment. Its drawback is its low sensitivity (in the order of picograms).

This method is usually used when it is necessary to quickly and easily analyze the presence or absence of a certain protein.

2.2 DETECTION BY CHEMIOLUMINISCENCE
In chemiluminescence assays, the enzyme bound to the secondary antibody triggers a reaction with a luminescent substrate generating light.

In this case, the great advantage is the high sensitivity provided by this method (in the order of femtograms), allowing proteins with very low levels of expression to be identified. As a drawback, note that it requires the use of specialized equipment to read the results.

3.- FLUORESCENT DETECTION METHODS
This type of detection is based on the use of secondary antibodies conjugated to fluorophores that produce signal by themselves, without the need to add any additional substrate.

Among the advantages, it should be noted that the signal is more stable than that produced by enzymatic detection methods and, above all, that the possibility of using fluorophorized antibodies with different emission wavelengths on the same Western Blot membrane allows multiplexing the experiments. It should also be noted that this method also allows quantifying the protein present in the sample.

As disadvantages, a lower sensitivity than chemiluminescence detection, and the need to use specialized equipment.

Fluorescence is among the most widely used Western Blot detection methods today.

ANTIBODIES FOR IMMUNOFLUORESCENCE

The immunofluorescence (IF) technique, based on the detection of a specific antigen of interest by using fluorescently labeled antibodies, is a technique widely used in research laboratories due to its simplicity and reliability.

The results can be visualized by fluorescence microscopy using short wavelengths and, in addition to detecting the presence or absence of a certain protein in the sample, it is possible to determine its distribution in the sample or confirm the presence of post-translational modifications, among others.

In this post we bring you some keys related to antibodies for immunofluorescence that can help you optimize the results of your tests.

3 KEYS WHEN USING ANTIBODIES FOR IMMUNOFLUORESCENCE

1.- THE IMPORTANCE OF THE SPECIFICITY OF ANTIBODIES FOR IMMUNOFLUORESCENCE
As in any other immunoassay, the specificity of the primary antibody against our target antigen is a determining factor in the reliability and success of the results. The more specific the antibody, the better the signal obtained and the less background noise generated.

Let us also remember that an antibody that has a high specificity against an antigen in a certain technique does not have to do so in another, even if it is the same antigen. Hence the importance of validating each antibody for each technique in which it will be used. In the case at hand, it is essential to previously validate the immunofluorescence antibodies to be used in the assay.

How can we validate the antibodies for immunofluorescence? For there are various methods such as positive and negative expression experiments using, for example, knock-out cell lines, by experimental manipulation of the location of the target protein, protocol optimizations, etc. Or, resorting to commercial antibodies already validated for use in this technique.

2.- CONTROLS FOR IMMUNOFLUORESCENCE
The inclusion of controls, as in any other experiment, will increase confidence in the results obtained in terms of specificity and sensitivity.

To avoid errors derived from autofluorescence phenomena or from nonspecific binding of antibodies, the use of negative controls in immunofluorescence assays is very important.

Additionally, additional controls such as omission of the primary antibody, the use of isotype controls and of negative and positive cell lines for the antigen of interest may be included.

3.- DILUTION OF THE ANTIBODIES FOR IMMUNOFLUORESCENCE
To optimize the results of the tests, another key point is the titration of the antibodies to determine the ideal dilution to use in each case. This will also vary depending on whether we are dealing with a purified antibody or an antiserum.

In this sense, it is important to achieve a good signal / background noise ratio, that is, an optimal relationship between the intensity of the fluorescent signal from the antigen of interest and the background signal due to nonspecific junctions. If we apply the primary antibody at a very low concentration, it will be very difficult to distinguish the positive signal. Conversely, an overly concentrated antibody will excessively increase background noise.

The typical concentration / dilution ranges for immunofluorescence experiments are usually between 1-10ug / mL in the case of using purified antibodies, and between 1: 100 – 1: 1000 for the antisera.

In this post you can remember some other recommended dilutions for other techniques and immunoassays.