What is the light chain for Human PSGL-1/CD162 Antibody, Catalog # MAB3345, Clone # 688102?

Name: Human PSGL-1/CD162 Antibody
Brand: R&D Systems
SKU: MAB3345
Price: 389 USD

TMAB3345 has a lambda light chain.

Human PSGL-1/CD162 Antibody

Catalog #: MAB3345 Datasheet / COA / SDS

Catalog #	Availability	Size / Price	Qty
MAB3345-100
MAB3345-SP

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All PSGL-1/CD162 Antibodies

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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 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

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.

Scientific Data

Immunohistochemistry

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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

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: 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

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