Key Product Details

Species Reactivity

Validated:

Mouse

Cited:

Human, Mouse, Transgenic Mouse, Xenograft

Applications

Validated:

Immunohistochemistry, Western Blot, Flow Cytometry, CyTOF-ready

Cited:

Immunohistochemistry, Immunohistochemistry-Paraffin, Immunohistochemistry-Frozen, Western Blot, Immunocytochemistry, ELISA Capture

Label

Unconjugated

Antibody Source

Polyclonal Goat IgG
View all PD-1 Primary Antibodies »

Product Specifications

Immunogen

S. frugiperda insect ovarian cell line Sf 21-derived recombinant mouse PD-1
Leu25-Gln167
Accession # Q02242

Specificity

Detects mouse PD-1 in direct ELISAs and Western blots. In direct ELISAs, approximately 20% cross-reactivity with recombinant human PD‑1 is observed.

Clonality

Polyclonal

Host

Goat

Isotype

IgG

Scientific Data Images for Mouse PD‑1 Antibody

Detection of Mouse PD-1 antibody by Western Blot.

Detection of Mouse PD‑1 by Western Blot.

Western blot shows lysates of EL-4 mouse lymphoblast cell line and CTLL-2 mouse cytotoxic T cell line (negative control). PVDF membrane was probed with 0.2 µg/mL of Goat Anti-Mouse PD-1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1021) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF017). A specific band was detected for PD-1 at approximately 55 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
Detection of Mouse PD-1 antibody by Western Blot.

Detection of Mouse PD‑1 by Western Blot.

Western blot shows lysates of 293T human embryonic kidney cell line mock transfected or transfected with mouse PD-1. PVDF membrane was probed with 0.2 µg/mL of Goat Anti-Mouse PD-1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1021) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF017). A specific band was detected for PD-1 at approximately 75 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
PD‑1 antibody in Mouse Thymus by Immunohistochemistry (IHC-Fr).

PD‑1 in Mouse Thymus.

PD‑1 was detected in perfusion fixed frozen sections of mouse thymus using 15 µg/mL Mouse PD‑1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1021) overnight at 4 °C. Tissue was stained (red) and counterstained (green). View our protocol for Fluorescent IHC Staining of Frozen Tissue Sections.
PD-1 antibody in Mouse Spleen by Immunohistochemistry (IHC-Fr).

PD‑1 in Mouse Spleen.

PD-1 was detected in perfusion fixed frozen sections of mouse spleen using Goat Anti-Mouse PD-1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1021) at 5 µg/mL overnight at 4 °C. Tissue was stained using the Anti-Goat HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS008) and counterstained with hematoxylin (blue). Specific staining was localized to cytoplasm in splenocytes. View our protocol for Chromogenic IHC Staining of Frozen Tissue Sections.
PD-1 antibody in Mouse Thymus by Immunohistochemistry (IHC-P).

PD‑1 in Mouse Thymus.

PD-1 was detected in perfusion fixed paraffin-embedded sections of mouse thymus using Goat Anti-Mouse PD-1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1021) at 1.7 µg/mL for 1 hour at room temperature followed by incubation with the Anti-Goat IgG VisUCyte™ HRP Polymer Antibody (Catalog # VC004). Before incubation with the primary antibody, tissue was subjected to heat-induced epitope retrieval using Antigen Retrieval Reagent-Basic (Catalog # CTS013). Tissue was stained using DAB (brown) and counterstained with hematoxylin (blue). Specific staining was localized to cell membranes. View our protocol for IHC Staining with VisUCyte HRP Polymer Detection Reagents.
Detection of Mouse PD-1 by Western Blot

Detection of Mouse PD-1 by Western Blot

Expression and glycosylation of Siglec-1 counter-receptors on resting and activated Tregs.(A) Eight Siglec-1 counter receptors were randomly selected for cytometry analysis. All experiments were performed three times and similar results were observed. (B) Normalized mRNA counts of Siglec-1 counter receptors were obtained from Treg RNA-Seq data. The result is visualized in a volcano plot. Only a small subset of counter-receptors showed strongly increased gene expression following Treg activation. The counter-receptors selected for flow cytometry analysis are highlighted in red. (C) Western blot showing that CD48 had a higher molecular weight in activated Tregs. (D) Western blot of PD-1 affinity-purified from resting and activated Tregs. PD 1 showed a dramatic decrease of molecular weight after PNGase F digestion (D), PD-1 from activated Tregs had higher molecular weight (E) and following sialidase treatment, PD-1 from resting and activated Tregs migrated at a slightly reduced molecular weight indicating the presence of sialic acids (F). Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/35224210), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of Mouse PD-1 by Western Blot

Detection of Mouse PD-1 by Western Blot

Expression and glycosylation of Siglec-1 counter-receptors on resting and activated Tregs.(A) Eight Siglec-1 counter receptors were randomly selected for cytometry analysis. All experiments were performed three times and similar results were observed. (B) Normalized mRNA counts of Siglec-1 counter receptors were obtained from Treg RNA-Seq data. The result is visualized in a volcano plot. Only a small subset of counter-receptors showed strongly increased gene expression following Treg activation. The counter-receptors selected for flow cytometry analysis are highlighted in red. (C) Western blot showing that CD48 had a higher molecular weight in activated Tregs. (D) Western blot of PD-1 affinity-purified from resting and activated Tregs. PD 1 showed a dramatic decrease of molecular weight after PNGase F digestion (D), PD-1 from activated Tregs had higher molecular weight (E) and following sialidase treatment, PD-1 from resting and activated Tregs migrated at a slightly reduced molecular weight indicating the presence of sialic acids (F). Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/35224210), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of Mouse PD-1 by Western Blot

Detection of Mouse PD-1 by Western Blot

Expression and glycosylation of Siglec-1 counter-receptors on resting and activated Tregs.(A) Eight Siglec-1 counter receptors were randomly selected for cytometry analysis. All experiments were performed three times and similar results were observed. (B) Normalized mRNA counts of Siglec-1 counter receptors were obtained from Treg RNA-Seq data. The result is visualized in a volcano plot. Only a small subset of counter-receptors showed strongly increased gene expression following Treg activation. The counter-receptors selected for flow cytometry analysis are highlighted in red. (C) Western blot showing that CD48 had a higher molecular weight in activated Tregs. (D) Western blot of PD-1 affinity-purified from resting and activated Tregs. PD 1 showed a dramatic decrease of molecular weight after PNGase F digestion (D), PD-1 from activated Tregs had higher molecular weight (E) and following sialidase treatment, PD-1 from resting and activated Tregs migrated at a slightly reduced molecular weight indicating the presence of sialic acids (F). Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/35224210), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of PD‑1 in HEK293 cell line transfected with mouse PD-1 and eGFP by flow cytometry.

HEK293 human embryonic kidney cell line transfected with (A) mouse PD-1 or (B) irrelevant transfected cell line and eGFP were stained with Goat Anti-Mouse PD‑1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1021) followed by Allophycocyanin-conjugated Anti-Goat IgG Secondary Antibody (Catalog # F0108). View our protocol for Staining Membrane-associated Proteins.

Applications for Mouse PD‑1 Antibody

Application
Recommended Usage

CyTOF-ready

Ready to be labeled using established conjugation methods. No BSA or other carrier proteins that could interfere with conjugation.

Flow Cytometry

0.25 µg/106 cells
Sample: HEK293 human embyonic kidney cell line transfected with mouse PD-1 and eGFP vs irrelevant HEK293 transfected cell line

Immunohistochemistry

5-15 µg/mL
Sample: Perfusion fixed frozen sections of mouse thymus and mouse spleen, and perfusion fixed paraffin-embedded sections of mouse thymus

Western Blot

0.2 µg/mL
Sample: EL‑4 mouse lymphoblast cell line and 293T human embryonic kidney cell line transfected with mouse PD-1

Reviewed Applications

Read 5 reviews rated 4.6 using AF1021 in the following applications:

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Flow Cytometry

Formulation, Preparation, and Storage

Purification

Antigen Affinity-purified

Reconstitution

Reconstitute at 0.2 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.

Formulation

Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. See Certificate of Analysis for details.
*Small pack size (-SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.

Shipping

Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.

Stability & Storage

Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 12 months from date of receipt, -20 to -70 °C as supplied.
  • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
  • 6 months, -20 to -70 °C under sterile conditions after reconstitution.

Calculators

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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Background: PD-1

Programmed Death-1 (PD-1) is type I transmembrane protein belonging to the CD28/CTLA-4 family of immunoreceptors that mediate signals for regulating immune responses (1). Other members of this family include CD28, CTLA-4, and ICOS (2-4). PD-1 is most closely related to CTLA-4 and shares approximately 24% amino acid (aa) sequence identity. The mouse PD-1 gene encodes a 288 aa protein with a putative 20 aa signal peptide, a 149 aa extracellular region with one immunoglobulin-like V-type domain, a 21 aa transmembrane domain, and a 98 aa cytoplasmic region. The cytoplasmic tail contains two tyrosine residues that form the immunoreceptor tyrosine-based inhibitory motif (ITIM) and immunoreceptor tyrosine-based switch motif (ITSM) that are important in mediating PD-1 signaling. Mouse and human PD-1 share approximately 69% aa sequence identity. Two B7 family proteins, PD-L1 (also called B7-H1) and PD-L2, have been identified as PD-1 ligands (5, 6). PD-1 is expressed on activated T cells, B cells, myeloid cells, and on a subset of thymocytes. PD-1 deficient mice have a defect in peripheral tolerance and spontaneously develop autoimmune diseases. Binding of PD-1 to PD-L1 or PD-L2 results in the inhibition of TCR-mediated proliferation and cytokine production as well as BCR-mediated signaling. PD-1 likely has an inhibitory role in regulating immune responses (1-4).

References

  1. Ishida, Y. et al. (1992) EMBO J. 11:3887.
  2. Sharpe, A.H. and G.J. Freeman (2002) Nat. Rev. Immunol. 2:116.
  3. Coyle, A. and J. Gutierrez-Ramos (2001) Nat. Immunol. 2:203.
  4. Nishimura, H. and T. Honjo (2001) Trends in Immunol. 22:265.
  5. Latchman Y. et al. (2001) Nature Immun. 2:261.
  6. Tamura, H. et al. (2001) Blood 97:1809.

Long Name

Programmed Death-1

Alternate Names

CD279, PD1, PDCD1, SLEB2

Entrez Gene IDs

5133 (Human); 18566 (Mouse); 301626 (Rat); 100533201 (Porcine); 486213 (Canine); 102123659 (Cynomolgus Monkey)

Gene Symbol

PDCD1

UniProt

Q02242

Additional PD-1 Products

Product Documents for Mouse PD‑1 Antibody

Certificate of Analysis

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Product Specific Notices for Mouse PD‑1 Antibody

For research use only

Citations for Mouse PD‑1 Antibody

Customer Reviews for Mouse PD‑1 Antibody (5)

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Showing  1 - 5 的 5 reviews Showing All
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  • Mouse PD-1 Antibody
    Name: Efstathios Stamatiades
    Application: Immunocytochemistry/Immunofluorescence
    Sample Tested: Mammary gland tissue
    Species: Mouse
    Verified Customer | Posted 07/09/2019
    Cryosections from PLP immersion fixed PyMT mammary gland tumors were stained for PD1 (1:50) for 4h @ RT. After washing, tissue was stained with Donkey anti-Goat alexa546 2ndary antibody for 1h at RT. CD3 in green, PD1 in red, nuclear staining (Hoechst) in blue
    Mouse PD‑1 Antibody AF1021
  • Mouse PD-1 Antibody
    Name: Ivan Luzardo
    Application: Western Blot
    Sample Tested: HCT-116 human colorectal carcinoma cell line
    Species: Human
    Verified Customer | Posted 03/28/2019
  • Mouse PD-1 Antibody
    Name: Anonymous
    Application: Functional Assay
    Sample Tested: mouse antibody
    Species: Mouse
    Verified Customer | Posted 03/07/2018
  • Mouse PD-1 Antibody
    Name: Anonymous
    Application: ELISA
    Sample Tested: Human recombinant antibody
    Species: Mouse
    Verified Customer | Posted 09/29/2017
  • Mouse PD-1 Antibody
    Name: Sachin Puri
    Application: Flow Cytometry
    Sample Tested: Melanoma tissue and Human T cell
    Species: Human
    Verified Customer | Posted 06/15/2017

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