Mouse PD-1 Biotinylated Antibody

Catalog #: BAF1021 Datasheet / COA / SDS

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BAF1021

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Mouse PD-1 Biotinylated Antibody Summary

Species Reactivity

Mouse

Specificity

Detects mouse PD-1 in Western blots. In Western blots, less than 5% cross-reactivity with recombinant mouse (rm) ICOS is observed and less than 1% cross-reactivity with rmCTLA-4 and rmCD28 is observed.

Source

Polyclonal Goat IgG

Purification

Antigen Affinity-purified

Immunogen

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

Formulation

Lyophilized from a 0.2 μm filtered solution in PBS with BSA as a carrier protein.

Label

Biotin

Applications

Recommended Concentration

Sample

Western Blot

0.1 µg/mL

Recombinant Mouse PD‑1 Fc Chimera (Catalog # 1021-PD)

Flow Cytometry

2.5 µg/10⁶ cells

Mouse CD4⁺ T cells treated with PHA

Immunohistochemistry

5-15 µg/mL

Immersion fixed paraffin-embedded sections of mouse spleen

Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.

Scientific Data

Immunohistochemistry

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PD‑1 in Mouse Spleen. PD‑1 was detected in immersion fixed paraffin-embedded sections of mouse spleen using Goat Anti-Mouse PD‑1 Biotinylated Antigen Affinity-purified Polyclonal Antibody (Catalog # BAF1021) at 10 µg/mL overnight at 4 °C. Before incubation with the primary antibody, tissue was subjected to heat-induced epitope retrieval using Antigen Retrieval Reagent-Basic (CTS013). Tissue was stained using the Anti-Goat HRP-DAB Cell & Tissue Staining Kit (CTS008) and counterstained with hematoxylin (blue). Specific staining was localized to lymphocytes. Staining was performed using our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.

Reconstitution Calculator

Preparation and Storage

Reconstitution

Reconstitute at 0.2 mg/mL in sterile PBS.

Shipping

The product is shipped at ambient temperature. Upon receipt, store it 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.

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