Granulysin Antibody [Unconjugated]

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 (AB-108-C, open histogram), followed by Phycoerythrin-conjugated Anti-Goat IgG Secondary Antibody (F0107).

Human PBMCs were stained with Rabbit Anti-Human NCAM‑1/CD56 PE‑conjugated Monoclonal Antibody (Catalog # FAB24086P) and either (A) Goat Anti-Human Granulysin Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3138) or (B) isotype control antibody (Catalog # AB-108-C) followed by Allophycocyanin-conjugated Anti-Goat IgG Secondary Antibody (Catalog # F0108). To facilitate intracellular staining, cells were fixed with Flow Cytometry Fixation Buffer (Catalog # FC004) and permeabilized with Flow Cytometry Permeabilization/Wash Buffer I (Catalog # FC005). View our protocol for Staining Intracellular Molecules.

• Reactivity: Hu
• Applications: WB, CyTOF-ready, Flow
• Clonality: Polyclonal
• Host: Goat
• Conjugate: Unconjugated
• Concentration: LYOPH

Granulysin Antibody [Unconjugated] Summary

• Immunogen: Mouse myeloma cell line NS0-derived recombinant human Granulysin
  Arg23-Leu145
  Accession # P22749
• 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: N/A
• Isotype: IgG
• Clonality: Polyclonal
• Host: Goat
• Gene: GNLY
• Purity Statement: Antigen Affinity-purified

Applications/Dilutions

• Dilutions:
  • CyTOF-ready
  • Intracellular Staining by Flow Cytometry 2.5 ug/10^6 cells
  • Western Blot 0.1 ug/mL

Packaging, Storage & Formulations

• 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.
• Buffer: Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. See Certificate of Analysis for details.
  *Small pack size (-SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.
• Preservative: No Preservative
• Concentration: LYOPH
• Reconstitution Instructions: Reconstitute at 0.2 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.

Notes

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for Granulysin Antibody [Unconjugated]

• 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

Background

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).

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.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Publications for Granulysin Antibody (AF3138)(2)

Publications using AF3138

• Ploeger, L;Kaleja, P;Tholey, A;Lettau, M;Janssen, O; Analysis of Cytotoxic Granules and Constitutively Produced Extracellular Vesicles from Large Granular Lymphocytic Leukemia Cell Lines Cells 2024-08-06 [PMID: 39195200] (Immunoprecipitation, Human)
• KR Brink, MG Hunt, AM Mu, K Groszman, KV Hoang, KP Lorch, BH Pogostin, JS Gunn, JJ Tabor An E. coli display method for characterization of peptide-sensor kinase interactions Nature Chemical Biology, 2022-12-08;0(0):. 2022-12-08 [PMID: 36482094] (Human)

Bioinformatics

• Gene Symbol: GNLY
• Uniprot:
  • P22749()