Human PSGL-1/CD162 Antibody

Catalog # Availability Size / Price Qty
AF3345
AF3345-SP
Human PSGL-1/CD162 Antibody in Data
3 Images
Product Details
Citations (2)
FAQs
Supplemental Products
Reviews

Human PSGL-1/CD162 Antibody Summary

Species Reactivity
Human
Specificity
Detects human PSGL-1/CD162 in direct ELISAs and Western blots.
Source
Polyclonal Sheep IgG
Purification
Antigen Affinity-purified
Immunogen
Chinese hamster ovary cell line CHO-derived recombinant human PSGL-1/CD162
Thr44-Val295
Accession # NP_002997
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.
Label
Unconjugated

Applications

Recommended Concentration
Sample
Western Blot
0.2 µg/mL
See below
ELISA

This antibody functions as an ELISA detection antibody when paired with Mouse Anti-Human PSGL‑1/CD162 Monoclonal Antibody(Catalog # MAB9962).

This product is intended for assay development on various assay platforms requiring antibody pairs. We recommend the Human PSGL-1/CD162 DuoSet ELISA (Catalog # DY3345-05) for convenient development of a sandwich ELISA.

 
Immunohistochemistry
3-15 µg/mL
See below

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 Examples

Western Blot Detection of Human PSGL-1/CD162 antibody by Western Blot. View Larger

Detection of Human PSGL‑1/CD162 by Western Blot. Western blot shows lysates of Jurkat human acute T cell leukemia cell line and human peripheral blood mononuclear cells (PBMCs). PVDF membrane was probed with 0.2 µg/mL of Sheep Anti-Human PSGL-1/CD162 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3345) followed by HRP-conjugated Anti-Sheep IgG Secondary Antibody (Catalog # HAF016). Specific bands were detected for PSGL-1/CD162 homodimer at approximately 250 kDa and PSGL-1/CD162 monomer at approximately 110 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 8.

Immunohistochemistry PSGL-1/CD162 antibody in Human Tonsil by Immunohistochemistry (IHC-P). View Larger

PSGL‑1/CD162 in Human Tonsil. PSGL-1/CD162 was detected in immersion fixed paraffin-embedded sections of human tonsil using Sheep Anti-Human PSGL-1/CD162 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3345) at 3 µg/mL overnight at 4 °C. Tissue was stained using the Anti-Sheep HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS019) and counterstained with hematoxylin (blue). Specific staining was localized to lymphocytes. View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.

Human PSGL‑1/CD162 ELISA Standard Curve. Recombinant Human PSGL-1/CD162 protein was serially diluted 2-fold and captured by Mouse Anti-Human PSGL-1/CD162 Monoclonal Antibody(Catalog # MAB9962) coated on a Clear Polystyrene Microplate (Catalog # DY990). Sheep Anti-Human PSGL-1/CD162 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3345) was biotinylated and incubated with the protein captured on the plate. Detection of the standard curve was achieved by incubating Streptavidin-HRP (Catalog # DY998) followed by Substrate Solution (Catalog # DY999) and stopping the enzymatic reaction with Stop Solution (Catalog # DY994).

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.

=
÷

Preparation and Storage

Reconstitution
Sterile PBS to a final concentration of 0.2 mg/mL.
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 (#16) and three tyrosines (#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 is 51%, 52% and 43% aa identical to 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

Product Datasheets

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.

2 Citations: Showing 1 - 2
Filter your results:

Filter by:

  1. Cell surface vimentin is an attachment receptor for enterovirus 71.
    Authors: Du, Ning, Cong, Haolong, Tian, Hongchao, Zhang, Hua, Zhang, Wenliang, Song, Lei, Tien, Po
    J Virol, 2014;88(10):5816-33.
    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;8(11):e78988.
    Species: Human
    Sample Types: Recombinant Protein
    Applications: Western Blot

FAQs

No product specific FAQs exist for this product, however you may

View all Antibody FAQs

Cell and Tissue Staining Kits

Immunohistochemistry Reagents

Isotype Controls

Reconstitution Buffers

Secondary Antibodies

Reviews for Human PSGL-1/CD162 Antibody

There are currently no reviews for this product. Be the first to review Human PSGL-1/CD162 Antibody and earn rewards!

Have you used Human PSGL-1/CD162 Antibody?

Submit a review and receive an Amazon gift card.

$25/€18/£15/$25CAN/¥75 Yuan/¥1250 Yen for a review with an image

$10/€7/£6/$10 CAD/¥70 Yuan/¥1110 Yen for a review without an image

Submit a Review