This protein datasheet indicates I need to use a cross-linking antibody, Catalog # MAB050, for biological activity. What is this antibody and is it really necessary?

Name: Recombinant Human Granulysin Protein, CF
Brand: R&D Systems
SKU: 3138-GN/CF

The antibody is directed against a 6x histidine repeat and is recommended for use as a cross-linker of proteins with 6x his-tag. Crosslinking is often used for proteins that require receptor trimerization and can result greater biological activity. R&D Systems Quality Control tests the performance of these proteins in the presence of the cross-linking antibody. Therefore, it is necessary to use this antibody when trying to achieve the same level of specific activity described in the datasheet.

Recombinant Human Granulysin Protein, CF

R&D Systems, Inc. a Bio-Techne Brand

Catalog # 3138-GN/CF

Citations (3)

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Source:	NS0
Accession #:	P22749
Applications:	Bioactivity

View Full Technical Details for Recombinant Human Granulysin Protein, CF

Product Specifications for Recombinant Human Granulysin Protein, CF

Source	Mouse myeloma cell line, NS0-derived human Granulysin protein Met1-Leu145, with a C-terminal 10-His tag
Purity	>95%, by SDS-PAGE under reducing conditions and visualized by silver stain
Endotoxin level	<1.0 EU per 1 μg of the protein by the LAL method.
N-terminal sequence Analysis	Arg23
Predicted Molecular Mass	15.4 kDa
SDS-PAGE	15 ‑ 16 kDa, reducing conditions
Activity	Measured by its ability to induce RANTES secretion by THP‑1 human acute monocytic leukemia cells. The ED₅₀ for this effect is 0.3-1 µg/mL, in the presence of 10 µg/mL of a cross-linking antibody Mouse Anti-polyHistidine Monoclonal Antibody (Catalog # MAB050).

Formulation, Preparation and Storage

Carrier Free

What does CF mean?

CF stands for Carrier Free (CF). We typically add Bovine Serum Albumin (BSA) as a carrier protein to our recombinant proteins. Adding a carrier protein enhances protein stability, increases shelf-life, and allows the recombinant protein to be stored at a more dilute concentration. The carrier free version does not contain BSA.

What formulation is right for me?

In general, we advise purchasing the recombinant protein with BSA for use in cell or tissue culture, or as an ELISA standard. In contrast, the carrier free protein is recommended for applications, in which the presence of BSA could interfere.

3138-GN/CF

Formulation	Lyophilized from a 0.2 μm filtered solution in Sodium Citrate and NaCl.
Reconstitution	Reconstitute at 100 μg/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. 3 months, -20 to -70 °C under sterile conditions after reconstitution.

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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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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).

Item	Value
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, LAG-2, LAG2, Lymphokine, Lymphokine LAG-2, NKG5, NKG5519, Protein NKG5, T-lymphocyte activation gene 519, TLA519, TLA519T-cell activation protein 519

Citations for Recombinant Human Granulysin Protein, CF (3)

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Citations are publications that use Bio-Techne products. Selected citations for Recombinant Human Granulysin Protein, CF include:

Species: Bacteria - E. Coli
Sample Types: Whole Cells
Applications: Bioassay

Hung-Mu Wei (2016), Antimicrobial Properties of an Immunomodulator - 15 kDa Human Granulysin PLoS ONE, 11(6):e0156321.
PMID: 27276051
Species: Yeast
Sample Types: Whole Cells
Applications: Bioassay

Longhi LN et al. (2012), Phenotypic and Functional Characterization of NK Cells in Human Immune Response against the Dimorphic Fungus Paracoccidioides brasiliensis. J. Immunol., 189(2):935-45.
PMID: 22723518
Species: Human
Sample Types: Whole Cells
Applications: Bioassay

Tewary P et al. (2010), Granulysin activates antigen-presenting cells through TLR4 and acts as an immune alarmin. Blood, 116(18):3465-74.
PMID: 20660289

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FAQs for Recombinant Human Granulysin Protein, CF

This protein datasheet indicates I need to use a cross-linking antibody, Catalog # MAB050, for biological activity. What is this antibody and is it really necessary?

The antibody is directed against a 6x histidine repeat and is recommended for use as a cross-linker of proteins with 6x his-tag. Crosslinking is often used for proteins that require receptor trimerization and can result greater biological activity. R&D Systems Quality Control tests the performance of these proteins in the presence of the cross-linking antibody. Therefore, it is necessary to use this antibody when trying to achieve the same level of specific activity described in the datasheet.

Recombinant Human Granulysin Protein, CF

R&D Systems, Inc. a Bio-Techne Brand

Key Product Details

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Product Specifications for Recombinant Human Granulysin Protein, CF

Formulation, Preparation and Storage

Carrier Free

Reconstitution Calculator

Background: Granulysin

Citations for Recombinant Human Granulysin Protein, CF (3)

FAQs for Recombinant Human Granulysin Protein, CF

Product Documents for Recombinant Human Granulysin Protein, CF

Product Datasheets

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