Human PSGL-1/CD162 Antibody

(2 citations)   
  • 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
  • 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
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 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.
Preparation and Storage
  • Reconstitution
    Sterile PBS to a final concentration of 0.2 mg/mL.
  • 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
Related Research Areas
Citations:

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
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Species
Applications
Sample Type
  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 Type: Whole Cells
    Application: Neut
  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 Type: Recombinant Protein
    Application: WB
Cell and Tissue Staining Kits
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Anti-Sheep HRP-DAB Cell & Tissue Staining Kit

CTS019 1
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Isotype Controls
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Normal Sheep IgG Control

Ctrl 5-001-A 9
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Secondary Antibodies
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Sheep IgG HRP-conjugated Antibody

WB, Simple Western HAF016 14  
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Donkey Anti-Sheep IgG NorthernLights™ NL557-conjugated Antibody

Flow, IHC, ICC NL010 3
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Donkey Anti-Sheep IgG NorthernLights™ NL637-conjugated Antibody

Flow, IHC, ICC NL011 5
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Donkey Anti-Sheep IgG NorthernLights™ NL493-conjugated Antibody

Flow, IHC, ICC NL012 2
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Sheep IgG (H+L) PE-conjugated Antibody

Flow F0126  
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Donkey Anti-Sheep IgG Biotinylated Antibody

WB BAF016 2
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Sheep IgG (H+L) APC-conjugated Antibody

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Sheep IgG VisUCyte HRP Polymer Antibody

IHC VC006  
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Donkey Anti-Sheep IgG (H+L) Fluorescein-conjugated Antibody

Flow F0125 1
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Donkey Anti-Sheep IgG (H+L) PerCP-conjugated Antibody

Flow F0128
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