Astana Biomed Group, an authorized Abcam distributor in Central Asia

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

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [NAT105] to PD1 - Chimeric
  • Suitable for: WB, IHC-P, Flow Cyt, IP
  • Reacts with: Human

Overview

  • Product name

    Anti-PD1 antibody [NAT105] - Chimeric
    See all PD1 primary antibodies

  • Description

    Rabbit monoclonal [NAT105] to PD1 - Chimeric

  • Host species

    Rabbit

  • Tested Applications & Species

    Application Species
    Flow Cyt
    Human
    IHC-P
    Human
    IP
    Human
    WB
    Human
    See all applications and species data

  • 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

Images

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

    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.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PD1 antibody [NAT105-EPR21106] - Chimeric (ab216352)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PD1 antibody [NAT105-EPR21106] - 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 (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).

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

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PD1 antibody [NAT105-EPR21106] - Chimeric (ab216352)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PD1 antibody [NAT105-EPR21106] - 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 (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).

  • Western blot - Anti-PD1 antibody [NAT105-EPR21106] - Chimeric (ab216352)
    Western blot - Anti-PD1 antibody [NAT105-EPR21106] - Chimeric (ab216352)
    Anti-PD1 antibody [NAT105] - 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).

  • Western blot - Anti-PD1 antibody [NAT105-EPR21106] - Chimeric (ab216352)
    Western blot - Anti-PD1 antibody [NAT105-EPR21106] - Chimeric (ab216352)
    All lanes : Anti-PD1 antibody [NAT105] - 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).

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

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