Human Granulysin Biotinylated Antibody

Catalog #: BAF3138 Datasheet / COA / SDS

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BAF3138

Detection of Granulysin in Human PBMCs by Flow Cytometry.

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Human Granulysin Biotinylated Antibody Summary

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.

Scientific Data

Intracellular Staining by Flow Cytometry

Detection of Granulysin antibody in Human PBMCs antibody by Flow Cytometry.

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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).

Reconstitution Calculator

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

Clayberger, C. and A.M. Krensky (2003) Curr. Opin. Immunol. 15:560.
Ma, L.L. et al. (2002) J. Immunol. 169:5787.
Wang, Z. et al. (2000) J. Immunol. 165:1486.
Linde, C.M.A. et al. (2005) Infect. Immun. 73:6332.
Yabe, T. et al. (1990) J. Exp. Med. 172:1159.
Hanson, D.A. et al. (1999) Mol. Immunol. 36:413.
Deng, A. et al. (2005) J. Immunol. 174:5243.
Walch, M. et al. (2005) J. Immunol. 174:4220.
Ernst, W.A. et al. (2000) J. Immunol. 165:7102.
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