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Rabbit Monoclonal PD1 antibody. Suitable for IHC-P, IP, WB, Flow Cyt (Intra) and reacts with Human samples.

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Images

Flow Cytometry (Intracellular) - Anti-PD1 antibody [NAT105] - Chimeric (AB216352), expandable thumbnail
  • Immunoprecipitation - Anti-PD1 antibody [NAT105] - Chimeric (AB216352), expandable thumbnail
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PD1 antibody [NAT105] - Chimeric (AB216352), expandable thumbnail
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PD1 antibody [NAT105] - Chimeric (AB216352), expandable thumbnail
  • Western blot - Anti-PD1 antibody [NAT105] - Chimeric (AB216352), expandable thumbnail

Key facts

Isotype

IgG

Host species

Rabbit

Storage buffer

pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA

Form

Liquid

Clonality

Monoclonal

Immunogen

  • The exact immunogen used to generate this antibody is proprietary information.

Reactivity data

Select an application
Product promiseTestedExpectedPredictedNot recommended
IHC-PIPWBFlow Cyt (Intra)
Human
Tested
Tested
Tested
Tested

Tested
Tested

Species

Human

Dilution info

1/50

Notes

Perform heat-mediated antigen retrieval before commencing with IHC staining protocol.

Tested
Tested

Species

Human

Dilution info

1/30

Notes

-

Tested
Tested

Species

Human

Dilution info

1/1000

Notes

-

Tested
Tested

Species

Human

Dilution info

1/600

Notes

-

Associated Products

Select an associated product type

9 products for Alternative Product

2 products for Alternative Version

Target data

Function

Inhibitory receptor on antigen activated T-cells that plays a critical role in induction and maintenance of immune tolerance to self (PubMed:21276005). Delivers inhibitory signals upon binding to ligands CD274/PDCD1L1 and CD273/PDCD1LG2 (PubMed:21276005). Following T-cell receptor (TCR) engagement, PDCD1 associates with CD3-TCR in the immunological synapse and directly inhibits T-cell activation (By similarity). Suppresses T-cell activation through the recruitment of PTPN11/SHP-2: following ligand-binding, PDCD1 is phosphorylated within the ITSM motif, leading to the recruitment of the protein tyrosine phosphatase PTPN11/SHP-2 that mediates dephosphorylation of key TCR proximal signaling molecules, such as ZAP70, PRKCQ/PKCtheta and CD247/CD3zeta (By similarity).The PDCD1-mediated inhibitory pathway is exploited by tumors to attenuate anti-tumor immunity and escape destruction by the immune system, thereby facilitating tumor survival (PubMed:28951311). The interaction with CD274/PDCD1L1 inhibits cytotoxic T lymphocytes (CTLs) effector function (PubMed:28951311). The blockage of the PDCD1-mediated pathway results in the reversal of the exhausted T-cell phenotype and the normalization of the anti-tumor response, providing a rationale for cancer immunotherapy (PubMed:22658127, PubMed:25034862, PubMed:25399552).

Alternative names

PD1, PD1, PDCD1, Programmed cell death protein 1, Protein PD-1, hPD-1

Recommended products

Rabbit Monoclonal PD1 antibody. Suitable for IHC-P, IP, WB, Flow Cyt (Intra) and reacts with Human samples.

Key facts

Isotype

IgG

Form

Liquid

Clonality

Monoclonal

Immunogen
  • The exact immunogen used to generate this antibody is proprietary information.
Clone number

NAT105

Purification technique

Affinity purification Protein A

Concentration
Loading...

Storage

Shipped at conditions

Blue Ice

Appropriate short-term storage duration

1-2 weeks

Appropriate short-term storage conditions

+4°C

Appropriate long-term storage conditions

-20°C

Aliquoting information

Upon delivery aliquot

Storage information

Avoid freeze / thaw cycle

Notes

This rabbit monoclonal chimeric antibody has been engineered from the mouse monoclonal parent antibody Anti-PD1 antibody [NAT105] (Anti-PD1 antibody [NAT105] ab52587). By necessity, some mouse sequence is retained as part of the variable domain.

We recommend using an Fc-directed, preadsorbed secondary antibody if multiplexing with mouse monoclonals. For example, Goat Anti-Rabbit IgG Fc (Alexa Fluor® 488) preadsorbed ab150097 (AF488), Goat Anti-Rabbit IgG Fc (Alexa Fluor® 594) preadsorbed ab150100 (AF594), Goat Anti-Rabbit IgG Fc (Alexa Fluor® 647) preadsorbed ab150099 (AF647) or Goat Anti-Rabbit IgG Fc (HRP) preadsorbed ab98467 (HRP).

Supplementary info

This supplementary information is collated from multiple sources and compiled automatically.

Activity summary

PD1 also known as Programmed Cell Death Protein 1 or PDCD1 is a transmembrane protein that plays a critical role in regulating immune responses. It has a mass of approximately 55 kDa. PD1 is expressed on the surface of T cells B cells and some myeloid cells. PD1’s expression increases upon activation of these immune cells assisting in maintaining peripheral tolerance. Researchers often use PD1 mouse models and chimeric antibodies to explore the function of PD1 for experimental purposes. Antibodies such as anti-PD1 such as EH12.2H7 help in blocking PD1 interaction to study its role further.

Biological function summary

PD1 serves as an inhibitory receptor acting as a checkpoint in the immune system. It becomes part of an immune-suppressive complex when it binds with its ligands PD-L1 or PD-L2 which are expressed on various cell types including some tumor cells. This interaction suppresses the proliferation of T cells and cytokine production contributing to immune homeostasis. By controlling T cell activity PD1 limits autoimmunity but can also reduce the immune system's capability to attack cancer cells.

Pathways

PD1 functions in the immune checkpoint pathway a critical regulatory circuit in immune regulation. The engagement of PD1 with its ligands initiates a cascade that inhibits the function and proliferation of T cells through downstream SHP-2 phosphatase activity. This pathway frequently involves other regulatory proteins like CTLA-4 and is an important mechanism by which the body modulates immune responses. Related pathways often intersect with those involving T cell receptor signaling and contribute to the overall modulation of immune activity.

Associated diseases and disorders

PD1 has a significant role in cancer and autoimmune disorders. PD1 expression can allow tumors to evade immune surveillance making PD1 a target for cancer therapies such as anti-PD1 antibodies which aim to block PD1 and restore T cell activity. The interaction of PD1 with cancer-related proteins like PD-L1 facilitates tumor immune evasion. In autoimmune disorders PD1’s regulation of immune balance can become dysregulated leading to persistent immune activation and tissue damage. Understanding PD1 and its interaction with proteins such as PD-L1 helps in developing therapeutic strategies for both cancer and autoimmune conditions.

Product promise

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6 product images

  • Flow Cytometry (Intracellular) - Anti-PD1 antibody [NAT105] - Chimeric (ab216352), expandable thumbnail

    Flow Cytometry (Intracellular) - Anti-PD1 antibody [NAT105] - Chimeric (ab216352)

    Intracellular flow cytometric analysis of MOLT-4 (humanlymphoblastic leukemia cell line) cell line treated with 500 ng/ml Ionomycin and 10 ng/ml Phorbol-12-myristate-13-acetate (PMA) for 24 hours (Red) or Untreated control (Green) labeling PD1 with ab216352 at 1/600 dilution compared with a Rabbit IgG, monoclonal [EPR25A] - Isotype Control (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) (black) and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (blue). Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) at 1/2000 dilution was used as the secondary antibody.

    Gated on viable cells.

  • Immunoprecipitation - Anti-PD1 antibody [NAT105] - Chimeric (ab216352), expandable thumbnail

    Immunoprecipitation - Anti-PD1 antibody [NAT105] - Chimeric (ab216352)

    PD1 was immunoprecipitated from 0.35 mg of MOLT-4 (human lymphoblastic leukemia cell line) treated with 500 ng/ml Ionomycin and 10 ng/ml Phorbol-12-myristate-13-acetate (PMA) for 24 hours whole cell lysate with ab216352 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab216352 at 1/500 dilution. VeriBlot for IP Detection Reagent (HRP) (VeriBlot for IP Detection Reagent (HRP) ab131366), was used for detection at 1/1,000 dilution

    Lane 1: MOLT-4 treated with 500 ng/ml Ionomycin and 10 ng/ml Phorbol-12-myristate-13-acetate (PMA) for 24 hours whole cell lysate 10 μg (Input).

    Lane 2: ab216352 IP in MOLT-4 treated with 500 ng/ml Ionomycin and 10 ng/ml Phorbol-12-myristate-13-acetate (PMA) for 24 hours whole cell lysate.

    Lane 3: Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of ab216352 in MOLT-4 treated with 500 ng/ml Ionomycin and 10 ng/ml Phorbol-12-myristate-13-acetate (PMA) for 24 hours whole cell lysate.

    Blocking and dilution buffer and concentration: 5% NFDM/TBST.

    Exposure time: 40 seconds.

    The expression profile observed is consistent with what has been described in the literature (PMID: 26318293).

    All lanes: Immunoprecipitation - Anti-PD1 antibody [NAT105] - Chimeric (ab216352)

    Predicted band size: 32 kDa

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PD1 antibody [NAT105] - Chimeric (ab216352), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PD1 antibody [NAT105] - Chimeric (ab216352)

    Immunohistochemical analysis of paraffin-embedded human tonsil tissue labeling PD1 with ab216352 at 1/50 dilution, followed by Rabbit specific IHC polymer detection kit HRP/DAB (Rabbit specific IHC polymer detection kit HRP/DAB ab209101) ready to use. Positive staining on T cells of human tonsil germinal center (PMID: 20087161; PMID: 17640856) is observed. Counter stained with hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Rabbit specific IHC polymer detection kit HRP/DAB (Rabbit specific IHC polymer detection kit HRP/DAB ab209101) ready to use.

    Perform antigen retrieval using Universal HIER antigen retrieval reagents (10X) (Universal HIER antigen retrieval reagent (10X) ab208572).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PD1 antibody [NAT105] - Chimeric (ab216352), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PD1 antibody [NAT105] - Chimeric (ab216352)

    Immunohistochemical analysis of paraffin-embedded human bladder carcinoma tissue labeling PD1 with ab216352 at 1/50 dilution, followed by Rabbit specific IHC polymer detection kit HRP/DAB (Rabbit specific IHC polymer detection kit HRP/DAB ab209101) ready to use. Positive staining on lymphocytes in human bladder carcinoma (PMID: 20087161, PMID: 17640856) is observed. Counter stained with hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Rabbit specific IHC polymer detection kit HRP/DAB (Rabbit specific IHC polymer detection kit HRP/DAB ab209101) Ready to use.

    Perform antigen retrieval using Universal HIER antigen retrieval reagents (10X) (Universal HIER antigen retrieval reagent (10X) ab208572).

  • Western blot - Anti-PD1 antibody [NAT105] - Chimeric (ab216352), expandable thumbnail

    Western blot - Anti-PD1 antibody [NAT105] - Chimeric (ab216352)

    Blocking/Dilution buffer: 5% NFDM/TBST.

    The expression profile observed is consistent with what has been described in the literature (PMID: 26318293).

    All lanes: Western blot - Anti-PD1 antibody [NAT105] - Chimeric (ab216352) at 1/1000 dilution

    All lanes: Human tonsil lysate at 10 µg

    Secondary

    All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/20000 dilution

    Developed using the ECL technique.

    Predicted band size: 32 kDa

    Observed band size: 50-55 kDa

    Exposure time: 3min

  • Western blot - Anti-PD1 antibody [NAT105] - Chimeric (ab216352), expandable thumbnail

    Western blot - Anti-PD1 antibody [NAT105] - Chimeric (ab216352)

    Blocking/Dilution buffer: 5% NFDM/TBST.

    The expression profile observed is consistent with what has been described in the literature (PMID: 26318293).

    All lanes: Western blot - Anti-PD1 antibody [NAT105] - Chimeric (ab216352) at 1/1000 dilution

    Lane 1: Untreated MOLT-4 (human lymphoblastic leukemia cell line) whole cell lysate at 10 µg

    Lane 2: MOLT-4 treated with 500 ng/ml Ionomycin and 10 ng/ml Phorbol-12-myristate-13-acetate (PMA) for 24 hours, whole cell lysate at 10 µg

    Secondary

    All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/20000 dilution

    Developed using the ECL technique.

    Predicted band size: 32 kDa

    Observed band size: 50-55 kDa

    Exposure time: 81s

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Product protocols

For this product, it's our understanding that no specific protocols are required. You can:

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