Recombinant
RabMAb

Anti-PD1 antibody [NAT105-EPR21106] - Chimeric (ab216352)

Overview

  • Product name
    Anti-PD1 antibody [NAT105-EPR21106] - Chimeric
    See all PD1 primary antibodies
  • Description
    Rabbit monoclonal [NAT105-EPR21106] to PD1 - Chimeric
  • Host species
    Rabbit
  • Tested applications
    Suitable for: WB, IHC-P, Flow Cyt, IPmore details
  • Species reactivity
    Reacts with: Human
  • Immunogen

    Tissue, cells or virus corresponding to Human PD1. Specifically, human YT cells (NK-like leukemia cell line) that express PD1.
    Database link: Q15116

  • Positive control
    • WB: MOLT-4 treated with 500 ng/ml Ionomycin and 10 ng/ml Phorbol-12-myristate-13-acetate (PMA) for 24 hours, whole cell lysate; Human tonsil lysate. IHC-P: Human tonsil and bladder carcinoma tissues. Flow Cyt: MOLT-4 cells treated with 500 ng/ml Ionomycin and 10 ng/ml Phorbol-12-myristate-13-acetate (PMA) for 24 hours. IP: MOLT-4 treated with 500 ng/ml Ionomycin and 10 ng/ml Phorbol-12-myristate-13-acetate (PMA) for 24 hours whole cell lysate.
  • General notes

    This rabbit antibody has been engineered from the mouse monoclonal parent antibody 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, ab150097 (AF488), ab150100 (AF594), ab150099 (AF647) or ab98467 (HRP). 

Properties

Applications

Our Abpromise guarantee covers the use of ab216352 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
WB 1/1000. Detects a band of approximately 50-55 kDa (predicted molecular weight: 32 kDa).
IHC-P 1/50. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
Flow Cyt 1/600.
IP 1/30.

Target

  • Function
    Possible cell death inducer, in association with other factors.
  • Involvement in disease
    Genetic variation in PDCD1 is associated with susceptibility to systemic lupus erythematosus type 2 (SLEB2) [MIM:605218]. Systemic lupus erythematosus is a chronic, inflammatory and often febrile multisystemic disorder of connective tissue. It affects principally the skin, joints, kidneys and serosal membranes. It is thought to represent a failure of the regulatory mechanisms of the autoimmune system.
  • Sequence similarities
    Contains 1 Ig-like V-type (immunoglobulin-like) domain.
  • Developmental stage
    Induced at programmed cell death.
  • Cellular localization
    Membrane.
  • Information by UniProt
  • Database links
  • Alternative names
    • CD279 antibody
    • CD279 antigen antibody
    • hPD 1 antibody
    • hPD l antibody
    • hPD-1 antibody
    • hSLE1 antibody
    • PD 1 antibody
    • PD-1 antibody
    • PD1 antibody
    • PDCD 1 antibody
    • PDCD1 antibody
    • PDCD1_HUMAN antibody
    • Programmed cell death 1 antibody
    • Programmed cell death 1 protein antibody
    • Programmed cell death protein 1 antibody
    • Protein PD 1 antibody
    • Protein PD-1 antibody
    • SLEB2 antibody
    • Systemic lupus erythematosus susceptibility 2 antibody
    see all

Images

  • Flow cytometric analysis of MOLT-4 (human lymphoblastic 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 (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) (ab150077) at 1/2000 dilution was used as the secondary antibody.

    Gated on viable cells.

  • 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 (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 (ab209101) ready to use.

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

  • 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 secondary antibody (HRP) (ab131366), was used as secondary antibody at 1/1000 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 (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).

  • 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 (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 (ab209101) Ready to use.

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

  • Anti-PD1 antibody [NAT105-EPR21106] - Chimeric (ab216352) at 1/1000 dilution + Human tonsil lysate at 10 µg

    Secondary
    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
    why is the actual band size different from the predicted?


    Exposure time: 3 minutes


    Blocking/Dilution buffer: 5% NFDM/TBST.

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

  • All lanes : Anti-PD1 antibody [NAT105-EPR21106] - Chimeric (ab216352) at 1/1000 dilution

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

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : 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 why is the actual band size different from the predicted?


    Exposure time: 81 seconds


    Blocking/Dilution buffer: 5% NFDM/TBST.

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

References

ab216352 has not yet been referenced specifically in any publications.

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Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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