Overview

  • Product name

    Anti-PD1 antibody [CAL20]
    See all PD1 primary antibodies
  • Description

    Rabbit monoclonal [CAL20] to PD1
  • Host species

    Rabbit
  • Tested applications

    Suitable for: IHC-P, WB, ICC/IFmore details
    Unsuitable for: Flow Cyt or IP
  • Species reactivity

    Reacts with: Human
  • Immunogen

    Synthetic peptide. This information is considered to be commercially sensitive.

  • Positive control

    • WB: MOLT-4 (treated with 10ng/ml PMA and 500ng/ml Ionomycin for 24h) whole cell lysate. ICC/IF: MOLT-4 cells (treated with 10ng/ml PMA and 500ng/ml Ionomycin for 24h). IHC-P: Human tonsil, lymph node and lung carcinoma tissues.

Properties

Applications

Our Abpromise guarantee covers the use of ab237728 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
IHC-P 1/500. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
WB 1/1000. Detects a band of approximately 50-55 kDa (predicted molecular weight: 32 kDa).
ICC/IF 1/50.
  • Application notes
    Is unsuitable for Flow Cyt or IP.
  • 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

    • Immunohistochemical analysis of paraffin-embeded human tonsil tissue labeling PD1 with ab237728 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) ready to use. Positive staining on the human tonsil.is observed. Counter stained with hematoxylin.

      Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) ready to use.

      Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 30 mins.

      The section was incubated with ab237728 for 15 mins at room temperature.

      The immunostaining was performed on a Leica Biosystems BOND® RX instrument.

    • Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized MOLT-4 (human lymphoblastic leukemia cell line) cells labeling PD1 with ab237728 at 1/50 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing membranous staining in MOLT-4 cells treated with PMA (10ng/ml 24h) and Ionomycin(500ng/ml 24h). The nuclear counter stain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) (ab195889) at 1/200 dilution (red).

      Secondary antibody only control: Used PBS instead of primary antibody, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution.

      Cells were either untreated, ot treated with treated with PMA (10ng/ml 24h) and Ionomycin (500ng/ml 24h).

    • All lanes : Anti-PD1 antibody [CAL20] (ab237728) at 1/1000 dilution

      Lane 1 : Untreated MOLT-4 (human lymphoblastic leukemia cell line), whole cell lysate
      Lane 2 : MOLT-4 (treated with 10ng/ml PMA and 500ng/ml Ionomycin for 24h), whole cell lysate

      Lysates/proteins at 20 µg per lane.

      Secondary
      All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

      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 and dilution buffer: 5% NFDM/TBST.

    • Formalin-fixed, paraffin-embedded human lymph node tissue stained for PD1 using ab237728 at 0.125 µg/ml dilution in immunohistochemical analysis.

    • Formalin-fixed, paraffin-embedded human lung carcinoma tissue stained for PD1 using ab237728 at 0.125 µg/ml dilution in immunohistochemical analysis.

    References

    ab237728 has not yet been referenced specifically in any publications.

    Customer reviews and Q&As

    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Human Tissue sections (tonsil)
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: Tris/EDTA pH 9.0
    Permeabilization
    No
    Specification
    tonsil
    Blocking step
    Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 15% · Temperature: RT°C
    Fixative
    10% NBF

    Abcam user community

    Verified customer

    Submitted Aug 12 2019

    Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
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