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Human PSGL-1/CD162 Antibody

Catalog #: MAB996
3 Citations Datasheet / COA / SDS
Catalog # Availability Size / Price Qty
MAB996
MAB996-SP
Product Details
Citations (3)
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Summary Preparation and Storage Reconstitution Calculator Background Product Datasheets Related Research Areas

Human PSGL-1/CD162 Antibody Summary

Species Reactivity
Human
Specificity
Detects human PSGL‑1/CD162. Recognizes sLex-bearing core 2 O‑glycan stuctures. It does not recognize sLex on an extended core 1 O‑glycan. The sLex-bearing, core 2 O‑glycan structure decorates the P-Selectin ligand PSGL-1, and the presence of this glycan structure is required for high affinity P-Selectin binding (1). This antibody stains human and canine leukocytes but does not recognize monkey, mouse, rabbit, porcine, feline or bovine leukocytes.
Source
Monoclonal Mouse IgM Clone # CHO131
Purification
IgM-specific Affinity-purified from hybridoma culture supernatant
Immunogen
CHO Chinese hamster ovary cell line transfected with human PSGL‑1/CD162
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.
Purity
IgM-specific Affinity-purified from hybridoma culture supernatant

Applications

Recommended Concentration
Sample
Flow Cytometry
2.5 µg/106 cells
Human whole blood monocytes and granulocytes
Adhesion Blockade
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 50 µg/mL of the antibody.
 
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.

Reconstitution Calculator

Reconstitution Calculator

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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Preparation and Storage

Reconstitution
Reconstitute at 0.5 mg/mL in sterile PBS.
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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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Citations for Human PSGL-1/CD162 Antibody

R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

3 Citations: Showing 1 - 3
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  1. Up-Regulated Expression of Matrix Metalloproteinases in Endothelial Cells Mediates Platelet Microvesicle-Induced Angiogenesis
    Authors: C Sun, SB Feng, ZW Cao, JJ Bei, Q Chen, WB Zhao, XJ Xu, Z Zhou, ZP Yu, HY Hu
    Cell. Physiol. Biochem., 2017-04-27;41(6):2319-2332.
    Species: Human
    Sample Types: Whole Cells
    Applications: Neutralization
  2. Bromelain decreases neutrophil interactions with P-selectin, but not E-selectin, in vitro by proteolytic cleavage of P-selectin glycoprotein ligand-1.
    Authors: Banks, Jessica, Herman, Christin, Bailey, Ryan C
    PLoS ONE, 2013-11-11;8(11):e78988.
    Species: Human
    Sample Types: Whole Cells
    Applications: Flow Cytometry
  3. P-selectin glycoprotein ligand-1 regulates chemokine-dependent leukocyte recruitment in colonic ischemia-reperfusion.
    Authors: Santen S, Schramm R, Menger MD, Wang Y, Jeppsson B, Thorlacius H
    Inflamm. Res., 2007-11-01;56(11):452-8.
    Species: Mouse
    Sample Types: In Vivo
    Applications: Neutralization

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