Product Information

Contents: Phycoerythrin (PE) anti-mouse Granzyme B
Catalog Number: 12-8822
Sizes: 25 ug
Formulation: Phosphate buffer pH 7.2,
150 mM NaCl, 0.09% NaN₃
Storage Conditions: Store at 4°C.
DO NOT FREEZE.
LIGHT-SENSITIVE MATERIAL.
Clone: 16G6
Isotype: Rat IgG2b, κ

C57BL/6 splenocytes were treated with 500 U/ml mouse IL-2 for 3 days and stained with FITC anti-mouse CD8a (53-6.7) (cat. 11-0081). Subsequently, cells were fixed and permeabilized (cat. 88-8823) and stained with PE Rat IgG2b Isotype Control (cat. 12-4031) (left) or 0.125 μg of PE 16G6 (right). Total cells were used for analysis.

Available Formats of This Product
Cat. No.	Format	Excite (nm)	Emit (nm)	Reported Applications
14-8822	Affinity Purified anti-mouse Granzyme B	N/A	N/A	ELISA cap WB
13-8822	Biotin anti-mouse Granzyme B	N/A	N/A	IC Flow WB
11-8822	Fluorescein isothiocyanate (FITC) anti-mouse Granzyme B	488	518	IC Flow
12-8822	Phycoerythrin (PE) anti-mouse Granzyme B	488	575	IC Flow
25-8822	Phycoerythrin-Cy7 (PE-Cy7) anti-mouse Granzyme B	488	760	IC Flow
51-8822	Alexa Fluor® 647 anti-mouse Granzyme B	633	668	IC Flow
Flow Cytometry Product Notes: Test Sizes: To accommodate multicolor flow cytometry, eBioscience is in the process of reducing test size volumes from 20 µl to 5 µl. Please check your antibody vial for the recommended test size. Fluorochrome Replacements: eBioscience is in the process of replacing all Pacific Blue® and APC-Alexa Fluor® 750 conjugated products with eFluor™ 450 and APC-eFluor™ 780 conjugated products, respectively.

Custom Product Requests
Need a custom product? Download the Custom Product Request Form and submit completed form to customs@ebioscience.com.

Questions? Please consult our answers to frequently asked questions at http://www.ebioscience.com/faq.

Description

The 16G6 antibody reacts with mouse Granzyme B (GrB) which is a member of the granzyme serine protease family. GrB is found in the granules of cytotoxic T cells and NK cells. Granzyme B has also been described as CGL1 (cathepsin G-like-1), a serine protease expressed only in cytotoxic T-lymphocytes after cell activation. GrB has been called CTLA-1 (cytotoxic T lymphocyte-associated serine esterase 1) based on identification of mRNA in various cytotoxic T cells, but not observed in non-cytotoxic lymphoid cells. GrB is crucial for the rapid induction of target cell death by apoptosis, induced by interaction with cytotoxic T cells. The receptor involved has been identified as mannose 6-phosphate receptor. This receptor functions as a death receptor for granzyme B during cytotoxic T cell-induced apoptosis.

For intracellular staining and flow cytometric analysis with direct conjugates of anti-mouse Granzyme B, it is highly recommended to use the Foxp3 Staining Buffer Set (cat. 00-5523). Other buffers may yield varying results. For more information, please contact technical support at tech@ebioscience.com.

Applications Reported

For research use only, not for diagnostic or therapeutic use. This 16G6 antibody has been reported for use in intracellular staining followed by flow cytometric analysis.

Applications Tested

This 16G6 antibody has been tested by intracellular staining and flow cytometric analysis of fixed and permeabilized IL-2 stimulated splenocytes using IC Cytokine Staining Protocol. This can be used at less than or equal to 0.125μg per million cells in 100 μl staining volume. It is recommended that the antibody be carefully titrated for optimal performance in the assay of interest.

References

Akha AA, Berger SB, Miller RA. Enhancement of CD8 T-cell function through modifying surface glycoproteins in young and old mice. Immunology. 2006 Oct;119(2):187-94. (16G6, Intracellular Flow, PubMed)

Kapp JA, Honjo K, Kapp LM, Xu XY, Cozier A, Bucy RP. TCR transgenic CD8+ T cells activated in the presence of TGFbeta express FoxP3 and mediate linked suppression of primary immune responses and cardiac allograft rejection. Int Immunol. 2006; 1-14. (16G6, IC flow, PubMed)

Kilinc MO, Aulakh KS, Nair RE, Jones SA, Alard P, Kosiewicz MM, Egilmez NK. Reversing Tumor Immune Suppression with Intratumoral IL-12: Activation of Tumor-Associated T Effector/Memory Cells, Induction of T Suppressor Apoptosis, and Infiltration of CD8+ T Effectors. J Immunol. 2006 Nov 15;177(10):6962-73 (16G6, IC Flow, PubMed)

Smyth, M., et al. 1995. Granzymes: exogenous proteinases that induce target cell apoptosis. Immunol Today. 16: 202-206.

Shafer-Weaver, K., et al. 2003. The Granzyme B ELISPOT assay: an alternative to the 51Cr-release assay for monitoring cell-mediated cytotoxicity. J. Translational Med. 1: 14.

Rininsland, F., et al. 2000. Granzyme B ELISPOT assay for ex vivo measurements of T cell immunity. J Immunol Meth. 240:143-155.

Related Products

Cat. 12-4031  Phycoerythrin (PE) Rat IgG2b Isotype Control
Cat. 88-8022  Mouse Granzyme B ELISA Ready-SET-Go!
Cat. 00-8222  eBioscience IC Fixation Buffer
Cat. 88-8322  Mouse Granzyme B ELISPOT Ready-SET-Go!
Cat. 00-8333  eBioscience Permeabilization Buffer(10X)
Cat. 88-8399  Human Granzyme B ELISPOT Ready-SET-Go!
Cat. 88-8823  eBioscience Fixation & Permeabilization
Cat. 12-8899  Phycoerythrin (PE) anti-human Granzyme B (clone GB11)
Cat. 14-8899  Affinity Purified anti-human Granzyme B (clone GB11)