Human Granulysin Biotinylated Antibody

  
  • Species Reactivity
    Human
  • Specificity
    Detects human Granulysin in Western blots. In Western blots, less than 1% cross-reactivity with recombinant human NKG-2D is observed.
  • Source
    Polyclonal Goat IgG
  • Purification
    Antigen Affinity-purified
  • Immunogen
    Mouse myeloma cell line NS0-derived recombinant human Granulysin
    Arg23-Leu145
    Accession # P22749
  • Formulation
    Lyophilized from a 0.2 μm filtered solution in PBS with BSA as a carrier protein.
  • Label
    Biotin
Applications
  •  
    Recommended
    Concentration
    Sample
  • Western Blot
    0.1 µg/mL
    Recombinant Human Granulysin
  • Intracellular Staining by Flow Cytometry
    2.5 µg/106 cells
    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
Intracellular Staining by Flow Cytometry
Detection of Granulysin in Human PBMCs by Flow Cytometry. Human peripheral blood mononuclear cells were treated for 24 hr with 50 ng/mL PMA and 500 ng/mL Ca2+ ionomycin, then stained with Goat Anti-Human Granulysin Biotinylated Antigen Affinity‑purified Polyclonal Antibody (Catalog # BAF3138, filled histogram) or control antibody (Catalog # BAF108, open histogram), followed by Streptavidin-Phycoerythrin (Catalog # F0040).
Preparation and Storage
  • Reconstitution
    Reconstitute at 0.2 mg/mL in sterile PBS.
  • Shipping
    The product is shipped at ambient temperature. Upon receipt, store it 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: Granulysin

Granulysin (formerly NKG5 or Lymphokine LAG-2) is a member of the saposin-like protein (SAPLIP) family of membrane disrupting proteins (1). Granulysin is expressed in granules of natural killer and activated cytotoxic T cells. It exhibits cytolytic activity against intracellular or extracellular microbes and also tumors, either alone or in synergy with perforin (2). Human granulysin has structural similarity and 30 - 40% aa identity to granulysins and NK-lysins of other mammals such as bovine, porcine and canine; similar peptides in rodents have not been identified (1). The 15 kDa unglycosylated protein contains five helical domains; helix 2 and 3 contain 9 arginines and one cysteine critical for activity. Peptides of either helix 2 or 3 will lyse bacteria, while helix 3 is needed to lyse tumor targets (3, 4). One isoform designated 519 uses a different start codon, has no signal peptide sequence and is poorly expressed (5). A post-translationally processed 9 kDa form is present in acidified granules and is less lytic than the 15 kDa form at granule pH (6). IL-15 is necessary and sufficient for granulysin upregulation in CD8 T cells (2). Nanomolar granulysin promotes chemotaxis and increases production of chemokines by monocytic cells, while micromolar local concentrations are needed for lysis (7). Experimental evidence supports the following mechanism for activity against intracellular pathogens (8). First, granulysin forms clusters in lipid rafts due to interaction of positive charges in helices 2-3 with acidic sphingolipids. After endocytosis, early endosomes fuse with phagosomes, probably regulated by small GTPase rab5. Granulysin binds microbial membranes through charge interactions and disrupts them, probably via scissoring actions of granulysin molecules (9, 10).

  • References:
    1. Clayberger, C. and A.M. Krensky (2003) Curr. Opin. Immunol. 15:560.
    2. Ma, L.L. et al. (2002) J. Immunol. 169:5787.
    3. Wang, Z. et al. (2000) J. Immunol. 165:1486.
    4. Linde, C.M.A. et al. (2005) Infect. Immun. 73:6332.
    5. Yabe, T. et al. (1990) J. Exp. Med. 172:1159.
    6. Hanson, D.A. et al. (1999) Mol. Immunol. 36:413.
    7. Deng, A. et al. (2005) J. Immunol. 174:5243.
    8. Walch, M. et al. (2005) J. Immunol. 174:4220.
    9. Ernst, W.A. et al. (2000) J. Immunol. 165:7102.
    10. Anderson, D.H. et al. (2003) J. Mol. Biol. 325:355.
  • Entrez Gene IDs:
    10578 (Human)
  • Alternate Names:
    D2S69Elymphocyte-activation gene 2; GNLY; Granulysin; LAG2; LAG-2; Lymphokine LAG-2; Lymphokine; NKG5; NKG5519; Protein NKG5; TLA519; TLA519T-cell activation protein 519; T-lymphocyte activation gene 519
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