PSGL-1/CD162 Antibody (688101) [Unconjugated]

Western blot shows lysates of Jurkat human acute T cell leukemia cell line. PVDF membrane was probed with 2 µg/mL of Mouse Anti-Human PSGL-1/CD162 Monoclonal Antibody (Catalog # MAB9961) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF018). Specific bands were detected for PSGL1/ CD162 homodimer at approximately 250 kDa and PSGL1/ CD162 monomer at approximately 110 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Human peripheral blood monocytes were stained with Mouse Anti-Human PSGL‑1/CD162 Monoclonal Antibody (Catalog # MAB9961, filled histogram) or isotype control antibody (Catalog # MAB003, open histogram), followed by Allophycocyanin-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # F0101B).

PSGL-1/CD162 was detected in immersion fixed human peripheral blood mononuclear cells (PBMCs) using Mouse Anti-Human PSGL-1/CD162 Monoclonal Antibody (Catalog # MAB9961) at 8 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Mouse IgG Secondary Antibody (red; Catalog # NL007) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm. View our protocol for Fluorescent ICC Staining of Non-adherent Cells.

PSGL‑1/CD162 was detected in immersion fixed paraffin-embedded sections of human tonsil using Mouse Anti-Human PSGL‑1/CD162 Monoclonal Antibody (Catalog # MAB9961) at 5 µg/mL for 1 hour at room temperature followed by incubation with the Anti-Mouse IgG VisUCyte™ HRP Polymer Antibody (Catalog # VC001). Tissue was stained using DAB (brown) and counterstained with hematoxylin (blue). Specific staining was localized to cytoplasm. View our protocol for IHC Staining with VisUCyte HRP Polymer Detection Reagents.

• Reactivity: Hu
• Applications: WB, Flow, IHC, CyTOF-ready, ICC/IF
• Clone: 688101
• Clonality: Monoclonal
• Host: Mouse
• Conjugate: Unconjugated
• Concentration: LYOPH

PSGL-1/CD162 Antibody (688101) [Unconjugated] Summary

• Immunogen: Chinese hamster ovary cell line CHO-derived recombinant human PSGL-1/CD162
  Gln42-Gly295
  Accession # NP_002997
• Specificity: Detects human PSGL-1/CD162 in direct ELISAs and Western blots.
• Source: N/A
• Isotype: IgG2a
• Clonality: Monoclonal
• Host: Mouse
• Gene: SELPLG
• Purity Statement: Protein A or G purified from hybridoma culture supernatant

Applications/Dilutions

• Dilutions:
  • CyTOF-ready
  • Flow Cytometry 2.5 ug/10^6 cells
  • Immunocytochemistry 8-25 ug/mL
  • Immunohistochemistry 5-25 ug/mL
  • Western Blot 2 ug/mL

Packaging, Storage & Formulations

• 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.
• Buffer: 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.
• Preservative: No Preservative
• Concentration: LYOPH
• Reconstitution Instructions: Sterile PBS to a final concentration of 0.5 mg/mL.

Notes

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for PSGL-1/CD162 Antibody (688101) [Unconjugated]

• CD162 antigen
• CD162
• P-selectin glycoprotein ligand 1
• PSGL1
• PSGL-1
• PSGL-1cutaneous lymphocyte-associated associated antigen
• selectin P ligandCLA
• SELPLG

Background

Human PSGL-1 (P-Selectin Glycoprotein Ligand-1; also CD162), is a 120 kDa mucin-type glycoprotein that plays a key role in leukocyte adhesion (1-3). It is synthesized as a 412 amino acid (aa) preproprecursor that contains a 17 aa signal sequence, a 24 aa propeptide, a 279 aa extracellular domain (ECD), a 21 aa transmembrane segment and a 71 aa cytoplasmic region (4, 5). Following cleavage of the pre- and prosegments, it is expressed as a 240 kDa disulfide-linked homodimer. The extreme N-terminus (aa 1-16 of the mature molecule) contains one threonine (aa 16) and three tyrosines (aa 5, 7, and 10) that are involved in ligand binding. The Thr residue allows for O-linked glycosylation in the form of a core-2 structure (GalNAc-Gal) linked in a beta 1,6 bond to a sialylated Lewis X motif (GlcNAc linked to both Fuc and Gal with a terminal sialic acid residue) (1, 2, 5, 6, 7). The three tyrosine residues allow for sulfation (8, 9). When binding to P-selectin, Tyr sulfation and glycosylation are essential. Tyr7 provides the most efficient sulfate moiety, while Fuc and sialic acid are essentially mandatory (7). When binding to E-selectin, only carbohydrate is needed, while both carbohydrate and Tyr10 are used for L-selectin binding (6, 8). There are 16 decameric aa repeats in the ECD of the longform of PSGL-1. This form is referred to as the A allele, and represents 65 - 80% of the population. Alleles B and C show deletions of decameric repeats #2 (aa 132-141) plus #9 and 10 (aa 222-241), respectively. Shorter forms may show weaker binding to P-selectin (9, 10). Soluble forms of PSGL-1 are also known. Neutrophil elastase will cleave somewhere within repeats #5-9, while cathepsin G cleaves after Tyr7 (11). The loss of Tyr5 and 7 should impact binding affinity. PSGL-1 is found on virtually all leukocytes and macrophages/DC’s (1). Although there is similarity in the organization of the ECD between species, there is little aa identity. Human PSGL-1 ECD shares 51%, 52% and 43% aa sequence identity with equine, canine and mouse ECD, respectively.

1. Yang, J. et al. (1999) Thromb. Haemost. 81:1.
2. Cummings, R.D. (1999) Braz. J. Med. Biol. Res. 32:519.
3. McEver, R.P. and R.D. Cummings (1997) J. Clin. Invest. 100:485.
4. Sako, D. et al. (1993) Cell 75:1179.
5. Veldman, G.M. et al. (1995) J. Biol. Chem. 270:16470.
6. Bernimoulin, M.P. et al. (2003) J. Biol. Chem. 278:37.
7. Leppanen, A. et al. (2000) J. Biol. Chem. 275:39569.
8. Sako, D. et al. (1995) Cell 83:323.
9. Afshar-Kharghan, V. et al. (2001) Blood 97:3306.
10. Lozano, M.L. et al. (2001) Br. J. Haematol. 115:969.
11. Gardiner, E.E. et al. (2001) Blood 98:1440.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Publications for PSGL-1/CD162 Antibody (MAB9961)(1)

Publication using MAB9961

• PG Coulon, S Roy, S Prakash, R Srivastava, N Dhanushkod, S Salazar, C Amezquita, L Nguyen, H Vahed, AM Nguyen, WR Warsi, C Ye, EA Carlos-Cru, UT Mai, L BenMohamed Upregulation of Multiple CD8+ T Cell Exhaustion Pathways Is Associated with Recurrent Ocular Herpes Simplex Virus Type 1 Infection J. Immunol., 2020-06-15;0(0):. 2020-06-15 [PMID: 32540992] (Flow Cytometry, Human)

Bioinformatics

• Gene Symbol: SELPLG
• Uniprot:
  • NP_002997()