Human PD-1 Antibody

Catalog #: MAB10868 Datasheet / COA / SDS

Catalog #	Availability	Size / Price	Qty
MAB10868-100
MAB10868-SP

Detection of PD-1 in HEK293 Human Cell Line Transfected with Human PD-1 and eGFP by Flow Cytometry

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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 # 1015829

Purification

Protein A or G purified from hybridoma culture supernatant

Immunogen

Human embryonic kidney cell HEK293-derived human PD-1
Leu25-Thr168
Accession # Q15116

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.

Endotoxin Level

<0.10 EU per 1 μg of the antibody by the LAL method.

Label

Unconjugated

Applications

Recommended Concentration

Sample

Flow Cytometry

0.25 µg/10⁶ cells

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

Blockade of Receptor-ligand Interaction

In a functional ELISA, 0.15-2.10 µg/mL of this antibody will block 50% of the binding of 5 µg/mL of Recombinant Human PD-L1/B7-H1 Fc Chimera (Catalog # 156-B7) to immobilized Recombinant Human PD‑1 His-tagged Protein (Catalog # 8986-PD) coated at 1 µg/mL (100 µL/well). At 10 µg/mL, this antibody will block >90% of the binding.

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

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 (A) human PD-1 or (B) irrelevant protein, and eGFP was stained with Mouse Anti-Human PD-1 Monoclonal Antibody (Catalog # MAB10868) followed by Allophycocyanin-conjugated Anti-Mouse IgG Secondary Antibody (F0101B). Quadrant markers were set based on Mouse IgG1 Isotype Control (MAB002). Staining was performed using our Staining Membrane-associated Proteins protocol.

Blockade of Receptor-ligand Interaction

View Larger

PD-L1/B7-H1 Binding to PD-1 is Blocked by Human PD-1 Antibody. In a functional ELISA, 0.15-2.10 µg/mL of this antibody will block 50% of the binding of 5 µg/mL of Recombinant Human PD-L1/B7-H1 Fc Chimera (156-B7) to immobilized Recombinant Human PD‑1 His-tagged Protein (8986-PD) coated at 1 µg/mL (100 µL/well). At 10 µg/mL, this antibody will block >90% of the binding.

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