Overview

  • Product name
    Anti-PD1 antibody [NAT105]
    See all PD1 primary antibodies
  • Description
    Mouse monoclonal [NAT105] to PD1
  • Host species
    Mouse
  • Specificity
    This antibody recognizes human PD-1, a checkpoint protein expressed by activated T and B cells. PD-1 is involved in the control of immune cell responses.
  • Tested applications
    Suitable for: WB, Flow Cyt, IHC-Fr, ICC/IF, IP, IHC-Pmore 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: human tonsil and YT cell line lysates. IHC: Human tonsil, follicular lymphoma, lymph node and spleen (normal). ICC/IF: human tonsil cells, YT cells, MOLT4 cells. Flow Cyt: MOLT4 cells
  • General notes

    Please note that PD-1 is expressed variably in different tissues and that optimisation may be required depending on the tissue used for the experiment.

    Western blot protocol advice:

    Due to low expression of PD-1, we recommend loading a high amount of sample (100 µg) to detect the band for PD-1. Human tonsil and YT cell line lysates are suitable positive controls.

    This antibody clone [NAT105] is manufactured by Abcam.

    AF488 conjugate available as ab220300
    AF647 conjugate available as ab220301
    PE conjugate available as ab220302

    If you require this antibody in a different buffer formulation or a different conjugate for your experiments, please contact orders@abcam.com or you can find further information here.

Properties

Applications

Our Abpromise guarantee covers the use of ab52587 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 1/50. Predicted molecular weight: 32 kDa.
Flow Cyt 1/100.

ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.

IHC-Fr Use at an assay dependent concentration.
ICC/IF Use a concentration of 5 - 10 µg/ml.

We recommend Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) (ab150117) secondary antibody.

IP Use at an assay dependent concentration.
IHC-P 1/50. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

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

  • IHC image of PD1 staining in normal human tonsil formalin fixed paraffin embedded tissue section*, performed on a Leica Bond™ system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab52587 at 5 µg/ml for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

    For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

    *Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre

  • Double immunofluorescence staining of CD3 (green) and PD1 (red) on paraffin embedded tonsil.
  • Formaldehyde-fixed, paraffin-embedded Xhuman follicular lymphoma tissue stained for PD1 with ab52587 at 1/100 dilution in immunohistochemical analysis.

    See Abreview

  • Anti-PD1 antibody [NAT105] (ab52587) at 1/50 dilution + YT cell line extracts

    Predicted band size: 32 kDa
    Observed band size: 47 kDa (why is the actual band size different from the predicted?)

  • Paraffin-embedded human tonsil section stained with ab52587 PD1 antibody at 1/50 dilution.
  • Sample: Human tonsil cell extract
    Dilution: ab52587 antibody was used as 1:200 in 1x106 cells/tube.
    Anti-CD4 antibody was used as 1:200 dilution
    Anti-CD8 antibody was used as 1:200 dilution
  • MOLT4 cells stained for PD-1 (colored green) using ab52587 in ICC/IF. Cells were fixed with 100% methanol (5 min) at room temperature and incubated with PBS containing 10% goat serum, 0.3 M glycine, 1% BSA and 0.1% Tween-20 for 1 hour at room temperature to permeabilize the cells and block non-specific protein-protein interactions. The cells were then incubated with the PD-1 antibody ab52587 at 10 µg/ml and ab6046 (Rabbit polyclonal to beta Tubulin - Loading Control) at 1 ug/ml overnight at +4°C. The secondary antibodies used were Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) (ab150117) secondary antibody used at 1 ug/ml (colored green) and Goat Anti-Rabbit IgG H&L (Alexa Fluor® 594) (ab150080) secondary antibody (pseudo-colored red) used at 2 ug/ml for 1 hour at room temperature. DAPI was used to stain the cell nuclei (colored blue) at a concentration of 1.43 µM for 1 hour at room temperature.

  • Overlay histogram showing MOLT4 cells stained with ab52587 (red line). Live cells were incubated in 1x PBS / 10% normal goat serum to block non-specific protein-protein interactions followed by the antibody (ab52587, 1/100 dilution) for 30 min at 4°C. The secondary antibody used was Alexa Fluor® 488 goat anti-mouse IgG (H&L) (ab150117) at 1/2000 dilution for 30 min at 4°C.

    A mouse IgG1 isotype control antibody  (ab170190) was used at the same concentration and conditions as the primary antibody (black line). Unlabelled sample (blue line) was also used as a control.
     
    Acquisition of >5,000 events were collected using a 50 mW Blue laser (488nm) and 530/30 bandpass filter.
  • Immunohistochemical analysis of human large and locally advanced breast cancers staining PD-1 using ab52587. (a) Low level of PD-1+ T cell infiltration; (b) high level of PD-1+ T cell infiltration. (Itu: intratumoral Str: stromal)

  • Double immunoenzymatic staining of Ki67 (brown) and PD1 (red) on paraffin embedded tonsil.
  • Tissue Sample: Human Tonsil
    ab52587 diluted 1:100
    Incubation time: 30 mins
  • Immunohistochemical analysis of frozen Human liver tissue labeling PD1 with ab52587 at 1/50 dilution.

    See Abreview

  • Immunohistochemical analysis of various soft-tissue sarcomas staining PD1 using ab52587. Arrows indicate PD1 positive lymphocytes.

References

This product has been referenced in:
  • Maruse Y  et al. Significant association of increased PD-L1 and PD-1 expression with nodal metastasis and a poor prognosis in oral squamous cell carcinoma. Int J Oral Maxillofac Surg N/A:N/A (2018). IHC-P ; Human . Read more (PubMed: 29395669) »
  • Balzano T  et al. The Cerebellum of Patients with Steatohepatitis Shows Lymphocyte Infiltration, Microglial Activation and Loss of Purkinje and Granular Neurons. Sci Rep 8:3004 (2018). Read more (PubMed: 29445232) »

See all 64 Publications for this product

Customer reviews and Q&As

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Leica ER2
Sample
Monkey Tissue sections (Lymphoid tisssues - spleen, tonsil and lymph node)
Specification
Lymphoid tisssues - spleen, tonsil and lymph node
Permeabilization
No
Fixative
Formaldehyde
Username

Abcam user community

Verified customer

Submitted Jun 13 2014

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunohistochemistry (Frozen sections)
Sample
Human Tissue sections (Liver)
Specification
Liver
Fixative
Acetone/Methanol 1:2
Permeabilization
Yes - TBS + 0,1% TritonX100
Blocking step
Serum as blocking agent for 10 minute(s) · Concentration: 10% · Temperature: 20°C
Username

Abcam user community

Verified customer

Submitted Dec 15 2010

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Human Tissue sections (Follicular Lymphoma, Lymph node)
Specification
Follicular Lymphoma, Lymph node
Fixative
Formaldehyde
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Citrate Buffer, pH6
Blocking step
Novolink Protein Block, Novocastra, Leica Microsystems Inc. as blocking agent for 10 minute(s) · Concentration: 100% · Temperature: 25°C
Username

Dr. Hajnalka Rajnai

Verified customer

Submitted Nov 30 2009

Application
Immunohistochemistry (PFA perfusion fixed frozen sections)
Sample
Sheep Tissue sections (lung)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: citrate buffer
Permeabilization
No
Specification
lung
Blocking step
Serum as blocking agent for 30 minute(s) · Concentration: 25% · Temperature: 22°C
Fixative
Formaldehyde
Username

Ms. Anna Karagianni

Verified customer

Submitted Aug 11 2017

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Human Tissue sections (Human lymph node)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Low pH buffer
Permeabilization
No
Specification
Human lymph node
Blocking step
Dako endogenouse peroxidase block as blocking agent for 5 minute(s) · Concentration: 100% · Temperature: RT°C
Fixative
Paraformaldehyde
Username

Abcam user community

Verified customer

Submitted Nov 18 2015

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Human Tissue sections (Prostate Cancer)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Citrate buffer pH 6.0
Permeabilization
No
Specification
Prostate Cancer
Blocking step
Serum as blocking agent for 15 minute(s) · Concentration: 10% · Temperature: 23°C
Fixative
Formaldehyde
Username

Abcam user community

Verified customer

Submitted Nov 12 2015

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Blocking step
Dako Flex peroxidase as blocking agent for 5 minute(s) · Concentration: 100% · Temperature: RT°C
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Low pH
Sample
Human Tissue sections (Malignant Melanoma)
Specification
Malignant Melanoma
Permeabilization
No
Fixative
Formaldehyde
Username

Abcam user community

Verified customer

Submitted Jan 06 2015

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: TRIS-EDTA Buffer ph 8,5
Sample
Human Tissue sections (tonsil)
Specification
tonsil
Fixative
Paraformaldehyde
Username

Mr. Rudolf Jung

Verified customer

Submitted Oct 22 2014

.

You may want to consider the following secondary antibodies:

https://www.abcam.com/donkey-fab2-polyclonal-secondary-antibody-to-mouse-igg-hl-alexa-fluorreg-488-pre-adsorbed-ab150101.html

https://www.abcam.com/donkey-fab2-po...

Read More

I can confirm that the epitope of ab52587 is extracellular, because in FACS experiments fresh, unfixed cells were used.

1-10 of 11 Abreviews or Q&A

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