Human PD-1 Antibody

Catalog #: MAB10866 Datasheet / COA / SDS

Catalog #	Availability	Size / Price	Qty
MAB10866-100
MAB10866-SP

Detection of Human PD‑1 by Western Blot.

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Human PD-1 Antibody Summary

Species Reactivity

Human

Specificity

Detects human PD-1 in direct ELISAs.

Source

Monoclonal Mouse IgG₁ Clone # 1026519

Purification

Protein A or G purified from cell culture supernatant

Immunogen

Synthetic peptide containing human PD-1

Formulation

Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.

Label

Unconjugated

Applications

Recommended Concentration

Sample

Western Blot

2 µg/mL

NS0 mouse myeloma cell line transfected with human PD-1

Flow Cytometry

0.25 µg/10⁶ cells

HEK293 Human Cell Line transfected with Human PD-1 and eGFP

CyTOF-ready

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

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

Western Blot

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Detection of Human PD‑1 by Western Blot. Western blot shows lysates of NS0 mouse myeloma cell line mock transfected or transfected with human PD-1. PVDF membrane was probed with 2 µg/mL of Mouse Anti-Human PD‑1 Monoclonal Antibody (Catalog # MAB10866) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (HAF018). A specific band was detected for PD‑1 at approximately 75 kDa (as indicated). GAPDH (2275-PC-100) is shown as a loading control.This experiment was conducted under reducing conditions and using Western Blot Buffer Group 1.

Flow Cytometry

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Detection of PD-1 in HEK293 Human Cell Line transfected with Human PD-1 and eGFP by Flow Cytometry HEK293 human embryonic kidney cell line transfected with either (A) human PD-1 or (B) irrelevant protein, and eGFP, was stained with Mouse anti-human PD-1 monoclonal antibody (Catalog # MAB10866) followed by Allophycocyanin-conjugated anti-Mouse IgG Secondary Antibody (F0101B). Quadrant markers were set based on control antibody staining (MAB002, data not shown). Staining was performed using our Staining Membrane-Associated Proteins protocol.

Reconstitution Calculator

Preparation and Storage

Reconstitution

Reconstitute at 0.5 mg/mL in sterile 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.

Background: PD-1

Programmed Death-1 receptor (PD-1), also known as CD279, is type I transmembrane protein belonging to the CD28 family of immune regulatory receptors (1). Other members of this family include CD28, CTLA-4, ICOS, and BTLA (2-5). Mature human PD-1 consists of a 148 amino acid (aa) extracellular region (ECD) with one immunoglobulin-like V-type domain, a 24 aa transmembrane domain, and a 95 aa cytoplasmic region. The human PD-1 ECD shares 65% aa sequence identity with the mouse PD-1 ECD. 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 for mediating PD-1 signaling. PD-1 acts as a monomeric receptor and interacts in a 1:1 stoichiometric ratio with its ligands PD-L1 (B7-H1) and PD-L2 (B7-DC) (6, 7). PD‑1 is expressed on activated T cells, B cells, monocytes, and dendritic cells while PD-L1 expression is constitutive on the same cells and also on nonhematopoietic cells such as lung endothelial cells and hepatocytes (8, 9). Ligation of PD-L1 with PD-1 induces co-inhibitory signals on T cells promoting their apoptosis, anergy, and functional exhaustion (10). Thus, the PD-1: PD-L1 interaction is a key regulator of the threshold of immune response and peripheral immune tolerance (11). Finally, blockade of the PD-1: PD-L1 interaction by either antibodies or genetic manipulation accelerates tumor eradication and shows potential for improving cancer immunotherapy (12, 13, 14).

References