Human Granulysin Antibody

Catalog # Availability Size / Price Qty
AF3138
AF3138-SP
Human Granulysin Antibody in Intracellular Staining by Flow Cytometry
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Human Granulysin Antibody Summary

Species Reactivity
Human
Specificity
Detects human Granulysin in direct ELISAs and Western blots. In direct ELISAs and 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 Trehalose. *Small pack size (SP) is supplied as a 0.2 µm filtered solution in PBS.
Label
Unconjugated

Applications

Recommended Concentration
Sample
Western Blot
0.1 µg/mL
Recombinant Human Granulysin (Catalog # 3138-GN/CF)
CyTOF-ready
Ready to be labeled using established conjugation methods. No BSA or other carrier proteins that could interfere with conjugation.
 
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 Example

Intracellular Staining by Flow Cytometry Detection of Granulysin in Human PBMCs by Flow Cytometry. View Larger

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 200 ng/ml Ca2+ ionomycin then stained with Goat Anti-Human Granulysin Antigen Affinity‑purified Polyclonal Antibody (Catalog # AF3138, filled histogram) or control antibody (Catalog # AB‑108‑C, open histogram), followed by Phycoerythrin-conjugated Anti-Goat IgG Secondary Antibody (Catalog # F0107).

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 0.2 mg/mL in sterile PBS.
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: 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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Isotype Controls

Reconstitution Buffers

Secondary Antibodies

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