Recombinant human monoclonal antibody expressed in CHO binding to human PDCD1. It is a fully human IgG4 monoclonal antibody for the treatment of cancer. It acts as an immunomodulator by blocking ligand activation of the programmed cell death 1 (PD-1) receptor on activated T cells.
Figure 1 Anti-PDCD1 Recombinant Antibody (TAB-770) in ELISA
ELISA analysis of TAB-770 was performed by coating with PD-1 Protein, Human, Recombinant (His Tag).
Figure 2 Anti-PDCD1 Recombinant Antibody (TAB-770) in WB
WB analysis of TAB-770 was performed by with PD-1 Protein, Human, Recombinant (His Tag).
Lane 1: TAB-770 antibody
Lane 2: Negative control
Lane 3: 100ng PDCD1 antigen
Lane 4: 200ng PDCD1 antigen
Lane 5: 400ng PDCD1 antigen
Figure 3 SDS-PAGE analysis of anti-PD-1 antibody (Cat# TAB-770)
The purified anti-PD-1 antibody (Cat# TAB-770) was loaded in SDS-PAGE gels in β-mercaptoethanol-reduced (Lane 1) and non-reduced (Lane 2) conditions. Gel stained for 30 minutes with Coomassie Blue. As a result of different β-mercaptoethanol-reduced proteins (Heavy chain and Light chain) migrate as about 50 kDa and 25 kDa, respectively.
Figure 4 SEC-HPLC analysis of anti-PD-1 antibody (Cat# TAB-770)
Size exclusion chromatography (SEC-HPLC) is a technique of purity analysis and macromolecular separation based on molecule size. The purity of anti-PD-1 antibody (Cat# TAB-770) was determined by this method with Agilent 1200 Series.
Figure 5 ELISA analysis of anti-PD-1 antibody (Cat# TAB-770)
ELISA analysis of anti-PD-1 antibody (Cat# TAB-770) was performed by coating with recombinant human PD-1 protein (His tag). Then blocked with BSA and incubated with anti-PD-1 antibodies. The HRP-conjugated goat anti-human IgG was used as a secondary antibody. Detection was performed using TMB substrate and stopped with sulfuric acid. The absorbances were read on a spectrophotometer at 450 nm.
Figure 6 Western blot analysis of anti-PD-1 antibody (Cat# TAB-770)
Western blot analysis of anti-PD-1 antibody (Cat# TAB-770) was performed by loading human PD-1 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 anti-PD-1 antibodies and HRP goat anti-human IgG was used as a secondary antibody (1/2000).
Lane 1: Reduced antigen (0.1 μg)
Lane 2: Reduced antigen (0.3 μg)
Lane 3: Reduced antigen (0.6 μg)
Figure 7 Dot blot analysis of anti-PD-1 antibody (Cat# TAB-770)
Dot blot analysis of anti-PD-1 antibody (Cat# TAB-770) was performed by coating with human PD-1 protein (His tag).
The anti-PD-1 antibody incubation concentration: 2 μg/mL.
HRP-conjugated goat anti-human IgG was used as a secondary antibody (1/2000).
Figure 8 Sensorgram fit of human PD-1 analyte-binding anti-PD-1 antibody ligand (Cat# TAB-770)
Dilute the antibody ligand (5 μg/mL) and antigen analyte with running buffer. The ligand was injected to sample channel (Fc2) at a flow rate of 10 μL/min to reach a capture level of about 400 RU. The analyte was injected to Fc1-Fc2 of the channel at a flow rate of 30 μL/min for an association phase of the corresponding time. The association and dissociation process were all handling in the running buffer. Repeat 6 cycles of analyte according to analyte concentrations in ascending order.
Figure 9 PD-L1 binding to PD-1 was blocked by anti-PD-1 antibody (Cat# TAB-770), IC₅₀ = 246.6 ng/mL.
In a functional ELISA, the recombinant human PD‑1 protein (His tag) was coated at 2 µg/mL, 100 µL/well. Anti-PD-1 antibody (Cat# TAB-770) were 3-fold serial diluted from 20 µg/mL, 100 µL/well. Coated PD-1 protein was incubated with the human PD-L1 mFc chimera (5 µg/mL) and anti-PD-1 antibody (Cat# TAB-770). The PD-L1 binding to PD-1 was detected by HRP-goat anti-mouse IgG secondary antibody. Human IgG4 antibody (Cat# MOB-0408ZL, Creative Biolabs) was used as an isotype control.
Figure 10 Blockade of PD-1/PD-L1 interaction by anti-PD-1 antibody (Cat# TAB-770) resulted in an increase of IFN-γ production.
PBMCs were stimulated with 2 μg/mL anti-CD3 antibody (Cat# TAB-0416CL, Creative Biolabs) and cultured for 144 hours with PD-1 antibody (Cat# TAB-770, Creative Biolabs). Human IgG4 antibody (Cat# MOB-0408ZL, Creative Biolabs) was used as an isotype control. After cultivation, the supernatant was collected and the IFN-γ secretion level was analyzed using a human IFN gamma ELISA kit. The assay was performed in three parallel wells.
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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CAT | Product Name | Application | Type |
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MOB-1201z | Mouse Anti-PDCD1 Recombinant Antibody (clone 27G11) | ELISA, FC, ICC, IF, IHC | Mouse IgG1 |
HPAB-0184-YC | Mouse Anti-PDCD1 Recombinant Antibody (HPAB-0184-YC) | FC, FuncS | Mouse IgG |
HPAB-0185-YC | Mouse Anti-PDCD1 Recombinant Antibody (HPAB-0185-YC) | FC, FuncS | Mouse IgG |
HPAB-0186-YC | Mouse Anti-PDCD1 Recombinant Antibody (HPAB-0186-YC) | ELISA, FC | Mouse IgG |
HPAB-0187-YC | Human Anti-PDCD1 Recombinant Antibody (HPAB-0187-YC) | ELISA, FC | Chimeric (mouse/human) IgG1, κ |
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(Creative Biolabs Cat# TAB-770, RRID: AB_3112008)
For Research Use Only. Not For Clinical Use.
For research use only. Not intended for any clinical use. No products from Creative Biolabs may be resold, modified for resale or used to manufacture commercial products without prior written approval from Creative Biolabs.
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