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    pd1-antibody-cal20-ab237728.pdf

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Cell Biology Apoptosis Receptors Death Receptors
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RecombinantRabMAb

Recombinant Anti-PD1 antibody [CAL20] (ab237728)

  • Datasheet
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Multiplex immunohistochemistry - Anti-PD1 antibody [CAL20] (ab237728)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PD1 antibody [CAL20] (ab237728)
  • Immunocytochemistry/ Immunofluorescence - Anti-PD1 antibody [CAL20] (ab237728)
  • Western blot - Anti-PD1 antibody [CAL20] (ab237728)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PD1 antibody [CAL20] (ab237728)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PD1 antibody [CAL20] (ab237728)
  • Anti-PD1 antibody [CAL20] (ab237728)

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

Conjugates logo Related conjugates and formulations

Carrier Free

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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 proprietary to Abcam and/or its suppliers.

  • 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. mIHC: Human tonsil tissue.

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.
  • Storage buffer

    pH: 7.2
    Preservative: 0.05% Sodium azide
    Constituent: PBS
  • Concentration information loading...
  • Purity

    Protein A purified
  • Purification notes

    Purity >99%
  • Clonality

    Monoclonal
  • Clone number

    CAL20
  • Isotype

    IgG
  • Research areas

    • Cell Biology
    • Apoptosis
    • Receptors
    • Death Receptors
    • Immunology
    • Adaptive Immunity
    • T Cells
    • CD
    • Cancer
    • Tumor immunology
    • CD markers
    • Cancer
    • Invasion/microenvironment
    • Apoptosis
    • Other
    • Immunology
    • Adaptive Immunity
    • Regulatory T Cells
    • Cancer
    • Cell Death
    • Apoptosis
    • Other

Associated products

  • Alternative Versions

    • Anti-PD1 antibody [CAL20] - BSA and Azide free (ab251613)
  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab97051)
  • Isotype control

    • Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
  • Recombinant Protein

    • Recombinant human PD1 protein (Active) (ab174035)
  • Related Products

    • Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (ab195889)
    • PD-L1 / PD1 Multiplex IHC-IF Antibody Panel (PD-L1, PD1, CD68, CD3, Ki67, panCK) (ab269812)

Applications

The Abpromise guarantee

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)
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.
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.
  • Target information above from: UniProt accession Q15116 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 5133 Human
    • Omim: 600244 Human
    • SwissProt: Q15116 Human
    • Unigene: 158297 Human
    • 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.

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

    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

    Datasheets and documents

    • Datasheet download

      Download

    References (1)

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

    ab237728 has been referenced in 1 publication.

    • Chen Y  et al. The frequency and inter-relationship of PD-L1 expression and tumour mutational burden across multiple types of advanced solid tumours in China. Exp Hematol Oncol 9:17 (2020). PubMed: 32775040

    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
    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"
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