What are the key features of clone 10-1074?

Clone 10-1074 is a recombinant Human antibody that exhibits unusually potent neutralization activity and broad reactivity against HIV Env, including newly transmitted viruses. It exists as a disulfide-linked dimer of the heavy and light chain hetero-dimer under non-reducing conditions.

How is this antibody expressed and purified?

This antibody is expressed in HEK 293 cells as a combination of a heavy chain and a light chain derived from anti-HIV env monoclonal antibodies. It is rigorously purified to achieve greater than 95% purity as determined by SDS-PAGE, suitable for Flow Cytometry and Functional Studies.

Human Anti-HIV Env Recombinant Antibody (clone 10-1074)

CAT#: PABL-508

Recombinant Human Antibody (10-1074) 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. This antibody exhibits unusual potent neutralization, including broad reactivity against newly transmitted viruses.

Tested Data Published Data

SDS-PAGE

Figure 1 Human Anti-HIV Env Recombinant Antibody (PABL-508) in SDS-PAGE

SDS-PAGE analysis of PABL-508 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-508 was greater than 95% as determined by SDS-PAGE.

SEC-HPLC

Figure 2 Human Anti-HIV Env Recombinant Antibody (PABL-508) in SEC-HPLC

The purity of PABL-508 was 99.4989% 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

Figure 3 Human Anti-HIV Env Recombinant Antibody (PABL-508) in WB

Western blot analysis of PABL-508 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-508 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)

Figure 4 Human Anti-HIV Env Recombinant Antibody (PABL-508) in DB

Dot Blot analysis of PABL-508 was performed by coating with recombinant HIV-1 [HIV-1/Clade B/C (CN54)] gp120 protein (His tag).

PABL-508 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-508) in ELISA

ELISA analysis of PABL-508 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-508. 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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Figure 1 Antibody binding at the surface of cells infected with two HIV-1 reference strains.

CEM-NKR cells infected with HIV-1 (NLAD8) were incubated with 15 μg/ml of anti-Env monoclonal antibodies. The levels of antibody bound on infected (Gag-high) and bystander (Gag-low and Gag-negative) cells were then evaluated by flow cytometry.

Bruel, T., Guivel-Benhassine, F., Lorin, V., Lortat-Jacob, H., Baleux, F., Bourdic, K., ... & Schwartz, O. (2017). Lack of ADCC breadth of human nonneutralizing anti-HIV-1 antibodies. Journal of virology, 91(8), e02440-16.

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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
10-1074

Applications
FC, FuncS

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
This antibody has been reported for use in Flow Cytometry and Function Assay.

Target

Alternative Names
ENV; gp160; envelope glycoprotein; Envelope surface glycoprotein gp160, precursor; hypothetical protein; Envelope surface glycoprotein gp120; Envelope transmembrane domain

Gene ID
155971

UniProt ID
P04578

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Potent Neutralization Activity

This recombinant antibody, clone 10-1074, demonstrated potent neutralization activity in our functional assays against HIV Env. It is reported to have broad reactivity, and our data supports this. It is an essential tool for our research into viral envelope glycoproteins and immune evasion mechanisms.

Excellent For Flow Cytometry

We used this antibody for Flow Cytometry to detect Env expression on the surface of HEK 293 cells. The signal was strong and specific. The product is well-characterized, composed of a specific heavy and light chain combination, which provides confidence in its consistency and performance.

Q&As

What are the key features of clone 10-1074?

A: Clone 10-1074 is a recombinant Human antibody that exhibits unusually potent neutralization activity and broad reactivity against HIV Env, including newly transmitted viruses. It exists as a disulfide-linked dimer of the heavy and light chain hetero-dimer under non-reducing conditions.
How is this antibody expressed and purified?

A: This antibody is expressed in HEK 293 cells as a combination of a heavy chain and a light chain derived from anti-HIV env monoclonal antibodies. It is rigorously purified to achieve greater than 95% purity as determined by SDS-PAGE, suitable for Flow Cytometry and Functional Studies.

View the frequently asked questions answered by Creative Biolabs Support.

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Downloadable Resources

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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.

Datasheet

Datasheet - PABL-508.pdf

MSDS

MSDS - PABL-508.pdf

COA

Certificate of Analysis Lookup

To download a Certificate of Analysis, please enter a lot number in the search box below. Note: Certificate of Analysis not available for kit components.

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