MABS2269

Anti-phospho-PD-1 (Tyr248) Antibody, clone 407.6G12

• Synonym(s): CD279, Programmed cell death protein 1, Protein PD-1, hPD-1
• UNSPSC Code: 12352203

Properties

• biological source: mouse
• Quality Level: 200
• antibody form: purified antibody
• antibody product type: primary antibodies
• clone: 407.6G12, monoclonal
• mol wt: calculated mol wt 31.65 kDa; observed mol wt ~55 kDa
• purified by: using protein G
• species reactivity: mouse, human
• species reactivity (predicted by homology): monkey
• packaging: antibody small pack of 100
• technique(s): flow cytometry: suitable; western blot: suitable
• isotype: IgG2aκ
• epitope sequence: C-terminal half
• Protein ID accession no.: NP_005009
• UniProt accession no.: Q15116
• storage temp.: 2-8°C
• Gene Information: human ... PDCD1(5133)

Description

Specificity

Clone 407.6G12 is a mouse monoclonal antibody that specifically detects PD-1 (CD279) phosphorylated on tyrosine 248.

Immunogen

KLH-conjugated linear peptide corresponding to 13 amino acids surrounding phospho-tyrosine 248 from the C-terminal half of human PD-1 (CD279).

Application

Quality Control Testing

Evaluated by Western Blotting in lysate from Jurkat cells treated with pervanadate (0.1 mM; 5 min.at 37C).

Western Blotting Analysis (WB): A 1:1000 dilution of this antibody detected PD-1 phosphorylated on tyrosine 248 in lysate from Jurkat cells treated with pervanadate (0.1 mM; 5 min. at 37C), but not in lysate from untreated Jurkat cells.

Tested Applications

Western Blotting Analysis: A representative lot detected PD-1 (CD279) in Western Blotting application (Bu, X., et al. (2021). Cancer Immunol Res. 9(12):1465-1475).

Flow Cytometry Analysis: A representative lot detected PD-1 (CD279) in Flow Cytometry application (Bu, X., et al. (2021). Cancer Immunol Res. 9(12):1465-1475).

Note: Actual optimal working dilutions must be determined by end user as specimens, and experimental conditions may vary with the end user.

Target description

Programmed cell death protein 1 (UniProt: Q15116; also known as Protein PD-1, hPD-1, CD279) is encoded by the PDCD1 (also known as PD1) gene (Gene ID: 5133) in human. PD-1 is a monomeric inhibitory cell surface receptor involved in the regulation of T-cell function during immunity and tolerance. PD-1 and its ligands, PD-L1 and PD-L2 play a key role in the maintenance of peripheral tolerance, a process by which the quiescence of autoreactive mature T cells is maintained. PD-1 is a single-pass type I membrane protein, synthesized with a signal peptide (aa 1-20), which is subsequently cleaved off. The mature form contains an extracellular domain (aa 24-170), a transmembrane domain (aa 171-191), and a cytoplasmic domain (aa 192-288). PD-1 also contains a single N-terminal immunoglobulin variable region (IgV) like domain, an ITIM motif (aa 221-226) and an ITSM motif ( aa 247-251). Tyrosine 248 falls in ITSM motif and located within the intracellular domain of PD-1 and, phosphorylation of PD-1 at Tyrosine 248 is a significant post-translational modification that influences its function. Src family kinases, such as Lck and Fyn, are involved in the initial phosphorylation of PD-1. Upon binding of PD-1 to its ligands (PD-L1 or PD-L2), phosphorylation at Tyr248 triggers downstream signaling pathways. PD-1 phosphorylation at Tyr248 promotes the recruitment of the protein tyrosine phosphatase PTPN11/SHP-2 that mediates de-phosphorylation of key TCR proximal signaling molecules, such as ZAP70, PRKCQ/PKC theta and CD247/CD3zeta. The phosphorylation at Tyr248 is crucial for the full inhibitory effect of PD-1 on T-cell activity. It contributes to the suppression of T-cell proliferation, cytokine production, and cytotoxic activity. Studies demonstrate that phosphorylation of the ITSM motif of PD-1 marks dysfunctional T cells that may be rescued with PD-1 blockade. Detection of phospho-PD-1 in TILs is a potential biomarker for PD-1 immunotherapy responses. (Ref.: Bu, X., et al. (2021). Cancer Immunol Res. 9(12):1465-1475).

Physical form

Purified mouse monoclonal antibody IgG2a in buffer containing 0.1 M Tris-Glycine (pH 7.4), 150 mM NaCl with 0.05% sodium azide.

Reconstitution

1.0 mg/mL. Please refer to guidance on suggested starting dilutions and/or titers per application and sample type.

Storage and Stability

Recommended storage: +2°C to +8°C.

Other Notes

Concentration: Please refer to the Certificate of Analysis for the lot-specific concentration.

Disclaimer

Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.

Safety Information

• Storage Class Code: 12 - Non Combustible Liquids
• WGK: WGK 1
• Flash Point(F): Not applicable
• Flash Point(C): Not applicable