Human PSGL-1/CD162 Antibody

Catalog # Availability Size / Price Qty
MAB3345-100
MAB3345-SP
Human PSGL-1/CD162 Antibody in Immunohistochemistry
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Human PSGL-1/CD162 Antibody Summary

Species Reactivity
Human
Specificity
Detects human PSGL-1 in direct ELISAs.
Source
Monoclonal Mouse IgG1 Clone # 688102
Purification
Protein A or G purified from hybridoma culture supernatant
Immunogen
Chinese hamster ovary cell line CHO-derived recombinant human PSGL-1
Gln42-Gly295
Accession # Q14242
Formulation
Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied 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
Immunohistochemistry
5-25 µg/mL
See below
Neutralization
The adhesion of U937 human histiocytic lymphoma cells (5 x 104 cells/well, 30 minute preincubation with the antibody) to immobilized Recombinant Human P-Selectin/CD62P (Catalog # ADP3, 10 µg/mL, 100 µL/well) was inhibited by 10 µg/mL of the antibody.

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.

Data Example

Immunohistochemistry PSGL-1/CD162  in Human Tonsil. View Larger

PSGL-1/CD162 in Human Tonsil. PSGL‑1/CD162 was detected in immersion fixed paraffin-embedded sections of human tonsil using Mouse Anti-Human PSGL‑1/CD162 Monoclonal Antibody (Catalog # MAB3345) at 5 µg/mL for 1 hour at room temperature followed by incubation with the Anti-Mouse IgG VisUCyte™ HRP Polymer Antibody (Catalog # VC001). Before incubation with the primary antibody, tissue was subjected to heat-induced epitope retrieval using Antigen Retrieval Reagent-Basic (Catalog # CTS013). Tissue was stained using DAB (brown) and counterstained with hematoxylin (blue). Specific staining was localized to cytoplasm in lymphocytes. View our protocol for IHC Staining with VisUCyte HRP Polymer Detection Reagents.

Reconstitution Calculator

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Preparation and Storage

Reconstitution
Reconstitute at 0.5 mg/mL in sterile PBS.
Reconstitution Buffer Available
Reconstitution Buffer 1 (PBS)
Catalog #
Availability
Size / Price
Qty
RB01
Shipping
The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C
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: PSGL-1/CD162

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.

References
  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.
Long Name
P-Selectin Glycoprotein Ligand 1
Entrez Gene IDs
6404 (Human); 20345 (Mouse); 363930 (Rat)
Alternate Names
CD162 antigen; CD162; P-selectin glycoprotein ligand 1; PSGL1; PSGL-1; PSGL-1cutaneous lymphocyte-associated associated antigen; selectin P ligandCLA; SELPLG

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