Human Anti-HIV Env Recombinant Antibody (clone PGT121)
CAT#: PABL-509
Recombinant Human Antibody (PGT121) is capable of binding to HIV env, expressed in HEK 293 cells. Expressed as the combination of a heavy chain (HC) containing VH from anti-HIV env mAb and CH1-3 region of human IgG1 and a light chain (LC) encoding VL from anti-HIV env proteins mAb and CL of human light chain. Exists as a disulfide linked dimer of the HC and LC hetero-dimer under non-reducing condition.
Published Data
Tested Data
Figure 1 ELISA analysis of PGT121 binding to monomeric gp120, which was extracted from lysed virus supernatants: (A) 92BR020, (C) 92RW020, (E) JR-FL E168K/N192A, (G) IAVI C22.
Antibodies were also tested for cell surface Env binding by flow cytometry: (B) 92BR020, (D) 92RW020, (F) JR-FL E168K/N192A, (H) IAVI C22. mAb 2G12 was included as a control (gray). ELISA values are reported as optical density at 405 nm (OD405) and flow cytometry values are reported as mean fluorescence intensity (MFI).
Sok, D., Laserson, U., Laserson, J., Liu, Y., Vigneault, F., Julien, J. P., ... & Mahan, A. (2013). The effects of somatic hypermutation on neutralization and binding in the PGT121 family of broadly neutralizing HIV antibodies. PLoS pathogens, 9(11), e1003754.
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SDS-PAGE
Figure 1 Human Anti-HIV Env Recombinant Antibody (PABL-509) in SDS-PAGE
SDS-PAGE analysis of PABL-509 in β-mercaptoethanol-reduced (Lane 1) and non-reduced (Lane 2) conditions. Gel stained for 30 minutes with Coomassie Blue. As a result, the different reduced proteins (Heavy chain and Light chain) migrate as about 50 kDa and 25 kDa, respectively.
The purity of PABL-509 was greater than 95% as determined by SDS-PAGE.
SEC-HPLC
Figure 2 Human Anti-HIV Env Recombinant Antibody (PABL-509) in SEC-HPLC
The purity of PABL-509 was 98.7602% as determined by SEC-HPLC.
Column: 3 µm, 7.8 x 300 nm
Mobile phase: 150 mM Sodium Phosphate Buffer, pH 7.0
Detection: UV 280 nm
Injection: 10 µl
WB
Figure 3 Human Anti-HIV Env Recombinant Antibody (PABL-509) in WB
Western blot analysis of PABL-509 was performed by loading recombinant HIV-1 [HIV-1/Clade B/C (CN54)] gp120 protein, (His tag) onto a 12% Tris-HCl polyacrylamide gel. Proteins were transferred to a NC membrane and blocked with 5% skim milk for at least one hour. Membranes were probed with PABL-509 and HRP conjugated Anti-Human IgG as a secondary antibody (1: 2000). Chemiluminescent detection was performed.
Lane 1: Reduced antigen (0.1 μg)
Lane 2: Reduced antigen (0.3 μg)
Lane 3: Reduced antigen (0.6 μg)
DB
Figure 4 Human Anti-HIV Env Recombinant Antibody (PABL-509) in DB
Dot Blot analysis of PABL-509 was performed by coating with recombinant HIV-1 [HIV-1/Clade B/C (CN54)] gp120 protein (His tag).
PABL-509 incubation concentration: 2 μg/mL.
HRP conjugated Anti-Human IgG as a secondary antibody (1: 2000)
ELISA
Figure 5 Human Anti-HIV Env Recombinant Antibody (PABL-509) in ELISA
ELISA analysis of PABL-509 was performed by coating with recombinant HIV-1 [HIV-1/Clade B/C (CN54)] gp120 protein, (His tag). Then blocked with BSA and incubated with PABL-509. The HRP conjugated Anti-Human IgG as a secondary antibody (1: 2000). Detection was performed using TMB substrate and stopped with sulfuric acid. The absorbances were read on a spectrophotometer at 450 nm.
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Specifications
- Immunogen
- HIV gp160; envelope glycoprotein
- Host Species
- Human
- Type
- Human IgG
- Specificity
- Tested positive against HIV-1 Env
- Species Reactivity
- HIV
- Clone
- PGT121
- Applications
- ELISA 1:1000-1:20000
Neut Assay-dependent
Product Property
- Purity
- >95% as determined by SDS-PAGE
- Concentration
- Please refer to the vial label for the specific concentration.
- Buffer
- PBS
- Preservative
- No preservatives
- Storage
- Centrifuge briefly prior to opening vial. Store at +4°C short term (1-2 weeks). Aliquot and store at -20°C long term. Avoid repeated freeze/thaw cycles.
Applications
- Application Notes
- Assay-dependent
Target
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(Creative Biolabs Cat# PABL-509, RRID: AB_3111661)
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Product Notes
This is a product of Creative Biolabs' Hi-Affi™ recombinant antibody portfolio, which has several benefits including:
• Increased sensitivity
• Confirmed specificity
• High repeatability
• Excellent batch-to-batch consistency
• Sustainable supply
• Animal-free production
See more details about Hi-Affi™ recombinant antibody benefits.
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COA
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