Recombinant

Recombinant Anti-PD1 antibody [CAL20] (ab237728)

Reviews (1) Submit a question

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [CAL20] to PD1
  • Suitable for: IHC-P, mIHC, WB, ICC/IF
  • Reacts with: Human

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, mIHC, 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.

  • General notes

    Reproducibility is key to advancing scientific discovery and accelerating scientists’ next breakthrough.

    Abcam is leading the way with our range of recombinant antibodies, knockout-validated antibodies and knockout cell lines, all of which support improved reproducibility.

    We are also planning to innovate the way in which we present recommended applications and species on our product datasheets, so that only applications & species that have been tested in our own labs, our suppliers or by selected trusted collaborators are covered by our Abpromise™ guarantee.

    In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.

    We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.

    Applications & species from publications and Abreviews that have not been tested in our own labs or in those of our suppliers are not covered by the Abpromise guarantee.

    Please check that this product meets your needs before purchasing. If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, as well as customer reviews and Q&As.

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.
mIHC Use at an assay dependent concentration.
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

      • Multiplex immunohistochemistry - Anti-PD1 antibody [CAL20] (ab237728)
        Multiplex immunohistochemistry - Anti-PD1 antibody [CAL20] (ab237728)

        Fluorescence multiplex immunohistochemical analysis of normal human tonsil tissue (formalin-fixed paraffin-embedded section).

        Merged staining of anti-PD1 (ab237728; orange; Opal™520), anti-PDL1 (ab237726; green; Opal™540), anti-CD68 (ab192847; yellow; Opal™570), anti-CD3 (ab16669; red; Opal™620), anti-Ki67 (ab16667; light blue; Opal™650) and anti-PanCK (ab7753; grey; Opal™690).

        The immunostaining was performed on a Leica Biosystems BOND® RX instrument with an Opal™ 7-color automation IHC kit (NEL821001KT, Akoya Biosciences®).

        The section was incubated in six rounds of staining; in the order of ab237728 (1/500 dilution), ab237726 (1/500 dilution), ab192847 (1/300 dilution), ab16669 (1/300 dilution), ab16667 (1/200 dilution) and ab7753 (1/200 dilution); each using a separate fluorescent tyramide signal amplification system.

        Sodium citrate antigen retrieval (Leica ER1, pH6.0, 30 minutes) was used in between rounds of tyramide signal amplification to remove the antibody from the previous round, to avoid any cross-reactivity.

        DAPI (dark blue) was used as a nuclear counter stain.

        Microscopy and pseudocoloring of individual Opal™ dyes was performed using a Vectra Polaris.

      • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PD1 antibody [CAL20] (ab237728)
        Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PD1 antibody [CAL20] (ab237728)

        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.

      • Immunocytochemistry/ Immunofluorescence - Anti-PD1 antibody [CAL20] (ab237728)
        Immunocytochemistry/ Immunofluorescence - Anti-PD1 antibody [CAL20] (ab237728)

        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).

      • Western blot - Anti-PD1 antibody [CAL20] (ab237728)
        Western blot - Anti-PD1 antibody [CAL20] (ab237728)
        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.

      • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PD1 antibody [CAL20] (ab237728)
        Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PD1 antibody [CAL20] (ab237728)

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

      • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PD1 antibody [CAL20] (ab237728)
        Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PD1 antibody [CAL20] (ab237728)

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

      Protocols

      To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.

      Click here to view the general protocols

      References (0)

      Publishing research using ab237728? Please let us know so that we can cite the reference in this datasheet.

      ab237728 has not yet been referenced specifically in any publications.

      Customer reviews and Q&As

      Show All Reviews Q&A
      Submit a review Submit a question

      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-PD1 antibody [CAL20]

       Excellent
      Abreviews
      abreview image
      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
      Read More

      Abcam user community

      Verified customer

      Submitted Aug 12 2019

      Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
      For licensing inquiries, please contact partnerships@abcam.com