Is the anti-PDCD1 recombinant antibody suitable for use in Western blotting?

Yes, the anti-PDCD1 recombinant antibody (TAB-H55) is suitable for use in Western blotting. It provides specific binding to PDCD1, allowing for reliable detection in Western blot assays.

What are the storage recommendations for the anti-PDCD1 recombinant antibody?

The recommended storage condition for the anti-PDCD1 recombinant antibody (TAB-H55) is at -20°C or lower. For short-term storage, it can be kept at 2-8°C. To ensure stability, avoid repeated freeze-thaw cycles.

Can the anti-PDCD1 recombinant antibody be used in immunoprecipitation assays?

Yes, the anti-PDCD1 recombinant antibody (TAB-H55) can be used in immunoprecipitation assays. It provides specific binding to PDCD1, enabling the successful precipitation of the target protein from complex mixtures.

Is the anti-PDCD1 recombinant antibody effective in ELISA applications?

Yes, the anti-PDCD1 recombinant antibody (TAB-H55) is effective in ELISA applications. It has been validated for use in such assays and provides reliable detection of PDCD1.

What is the optimal dilution for using the anti-PDCD1 recombinant antibody in immunofluorescence?

The optimal dilution for using the anti-PDCD1 recombinant antibody (TAB-H55) in immunofluorescence is typically 1:100 to 1:500. It is advisable to perform a dilution series to determine the best working concentration for your specific experimental conditions.

Human Anti-PDCD1 Recombinant Antibody (TAB-H55)

CAT#: TAB-H55

This product is a recombinant Human antibody that recognizes PDCD1. The antibody was expressed in mammalian cells with chemically defined culture media and was purified by affinity chromatography.

Tested Data Gene Expression

Figure 1 Anti-PDCD1 Antibody (TAB-H55) in WB

Western blot analysis of TAB-H55 was performed with Recombinant Human PD1 Protein.
Goat Anti-Human IgG-HRP as a secondary antibody.
Lane 1: Reducing antigen (0.1 μg)
Lane 2: Reducing antigen (0.3 μg)
Lane 3: Reducing antigen (0.6 μg)

ELISA

Figure 2 Anti-PDCD1 Antibody (TAB-H55) in ELISA

ELISA analysis of TAB-H55 was performed by coating with Recombinant Human PD1 Protein.
The Goat Anti-Human IgG-HRP as a secondary antibody.

Figure 3 Anti-PDCD1 Antibody (TAB-H55) in DB

Dot blot analysis of TAB-H55 was performed with PD-1 Protein, Human, Recombinant (His Tag).
TAB-H55 incubation concentration: 2ng/μL.
NC: Negative control PC: Positive control

SDS-PAGE

Figure 4 SDS-PAGE analysis of anti-PD-1 antibody (Cat# TAB-H55)

The purified anti-PD-1 antibody (Cat# TAB-H55) 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.

SEC-HPLC

Figure 5 SEC-HPLC analysis of anti-PD-1 antibody (Cat# TAB-H55)

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-H55) was determined by this method with Agilent 1200 Series.

ELISA

Figure 6 ELISA analysis of anti-PD-1 antibody (Cat# TAB-H55)

ELISA analysis of anti-PD-1 antibody (Cat# TAB-H55) 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 7 Western blot analysis of anti-PD-1 antibody (Cat# TAB-H55)

Western blot analysis of anti-PD-1 antibody (Cat# TAB-H55) 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 8 Dot blot analysis of anti-PD-1 antibody (Cat# TAB-H55)

Dot blot analysis of anti-PD-1 antibody (Cat# TAB-H55) 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).

SPR

Figure 9 Sensorgram fit of human PD-1 analyte-binding anti-PD-1 antibody ligand (Cat# TAB-H55)

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.

Block

Figure 10 PD-L1 binding to PD-1 was blocked by anti-PD-1 antibody (Cat# TAB-H55), IC₅₀ = 442.9 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-H55) 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-H55). 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.

Cell-based Bioassay

Figure 11 Blockade of PD-1/PD-L1 interaction by anti-PD-1 antibody (Cat# TAB-H55) 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-H55, 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.

❮

❯

Normal Tissue Figure 1 Lymph node

Germinal center cells
Staining:

Medium

Intensity: Strong
Quantity: <25%
Location: Cytoplasmic/
membranous

* Image credit: Image credit: Human Protein Atlas https://v21.proteinatlas.org/images/35981/161493_A_9_8.jpg

Normal Tissue Figure 2 Tonsil

Germinal center cells
Staining:

High

Intensity: Strong
Quantity: 75%-25%
Location: Cytoplasmic/
membranous
Non-germinal center cells
Staining:

Medium

Intensity: Strong
Quantity: <25%
Location: Cytoplasmic/
membranous

* Image credit: Image credit: Human Protein Atlas https://v21.proteinatlas.org/images/35981/161493_A_6_8.jpg

RNA Expression Figure 3 RNA cell line category: Cell line enhanced (JURKAT, MOLT-4, RPMI-8226)

Cell lines ordered by descending RNA expression order.

* Image credit: Image credit: Human Protein Atlas https://v21.proteinatlas.org/ENSG00000188389-PDCD1

❮

❯

Specifications

Host Species
Human

Derivation
Humanized (from mouse)

Type
IgG4, κ

Specificity
PDCD1(programmed cell death 1, PD-1, PD1, CD279) [Homo sapiens]

Species Reactivity
Human

Applications
Inhib, Agonist

Related Disease
Solid tumors

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.

Target

Alternative Names
PDCD1;programmed cell death 1;programmed cell death protein 1;CD279;PD1;protein PD-1;PD-1;SLEB2;hPD-1;hPD-l;

Gene ID
5133

UniProt ID
Q15116

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Customer Review

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Breast cancer biomarkers at key points during disease progression

Michael Thompson
29.Mar,23

Great for PDCD1 Research

The Anti-PDCD1 Recombinant Antibody has been a fantastic addition to our PDCD1 research. Its specificity and sensitivity are outstanding, providing clear and reliable results. This antibody has become essential for our experiments, improving the accuracy and reliability of our assays.

Sarah Harris
22.Jun,22

Effective and Reliable

We incorporated this antibody into our PDCD1 assays, and it has been a great asset. The specificity for PDCD1 is precise, allowing for accurate detection and analysis. The reliable performance has made it an invaluable tool in our lab.

Brian White
30.Aug,21

Consistent High-Quality Results

This antibody has proven to be consistent and reliable in our assays. The high-quality, reproducible results have significantly improved our experiments, making it a crucial part of our research toolkit. The strong affinity and specificity for PDCD1 have been particularly beneficial.

Q&As

Is the anti-PDCD1 recombinant antibody suitable for use in Western blotting?

A: Yes, the anti-PDCD1 recombinant antibody (TAB-H55) is suitable for use in Western blotting. It provides specific binding to PDCD1, allowing for reliable detection in Western blot assays.
What are the storage recommendations for the anti-PDCD1 recombinant antibody?

A: The recommended storage condition for the anti-PDCD1 recombinant antibody (TAB-H55) is at -20°C or lower. For short-term storage, it can be kept at 2-8°C. To ensure stability, avoid repeated freeze-thaw cycles.
Can the anti-PDCD1 recombinant antibody be used in immunoprecipitation assays?

A: Yes, the anti-PDCD1 recombinant antibody (TAB-H55) can be used in immunoprecipitation assays. It provides specific binding to PDCD1, enabling the successful precipitation of the target protein from complex mixtures.
Is the anti-PDCD1 recombinant antibody effective in ELISA applications?

A: Yes, the anti-PDCD1 recombinant antibody (TAB-H55) is effective in ELISA applications. It has been validated for use in such assays and provides reliable detection of PDCD1.
What is the optimal dilution for using the anti-PDCD1 recombinant antibody in immunofluorescence?

A: The optimal dilution for using the anti-PDCD1 recombinant antibody (TAB-H55) in immunofluorescence is typically 1:100 to 1:500. It is advisable to perform a dilution series to determine the best working concentration for your specific experimental conditions.

View the frequently asked questions answered by Creative Biolabs Support.

Citations

Wiese, Teresa. Pharmacological targeting of acid sphingomyelinase increases CD4\(^+\) Foxp3\(^+\) regulatory T cell subsets in patients with major depression. Diss. Universität Würzburg, 2022. https://doi.org/10.25972/OPUS-23347

The study investigates the pharmacological inhibition of acid sphingomyelinase (ASM) and its effects on human T cells, both in vitro and in vivo. The research aimed to analyze how inhibiting ASM affects the immune response, particularly focusing on the role of CD4+ Foxp3+ regulatory T cells (Tregs). By understanding the sphingolipid metabolism in these cells, the study explores how changes in sphingolipid content and functionality can influence immune cell behavior, which has implications for treating autoimmune and inflammatory diseases, as well as viral infections .
Creative Biolabs significantly contributed to this research by providing essential reagents. They supplied the PD-1 inhibiting monoclonal antibody pembrolizumab and the CTLA-4 inhibiting monoclonal antibody tremelimumab (CAT#: TAB-H55). These reagents were crucial for the experiments involving the modulation of immune responses, allowing the researchers to study the effects of blocking specific immune checkpoints. The use of these antibodies enabled detailed examination of the underlying mechanisms affected by ASM inhibition, thus advancing the understanding of T cell regulation and the potential therapeutic applications .

Cite This Product

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(Creative Biolabs Cat# TAB-H55, RRID: AB_3112038)

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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 - TAB-H55.pdf

MSDS

MSDS - TAB-H55.pdf

COA

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