50-9774-41

antibody from Invitrogen Antibodies

Targeting: GZMM LMET1, MET1

Flow cytometry (Supportive data in Antibodypedia)

Antibody Data

Product number

50-9774-41

Provider

Invitrogen Antibodies

Product name

Anti-Granzyme M Monoclonal Antibody (4B2G4), eFluor 660, eBioscience™

Antibody type

Monoclonal

Antigen

Other

Description

Description: Granzyme M is one of five granzyme serine proteases that have been identified in humans. These proteins are expressed in the granules of NK cells and cytotoxic T cells, and are critical for the induction of target cell apoptosis through the cleavage of intracellular substrates. Although not as well characterized as some of the other family members, Granzyme M has been found to exhibit some distinct functions. These include the non-cytotoxic inhibition of cytomegalovirus replication, induction of apoptosis in tumor cells, and contribution to the TLR4-induced inflammatory process. Applications Reported: This 4B2G4 antibody has been reported for use in intracellular staining followed by flow cytometric analysis. Applications Tested: This 4B2G4 antibody has been pre-titrated and tested by intracellular staining followed by flow cytometric analysis of normal human peripheral blood cells using the Intracellular Fixation & Permeabilization Buffer Set (cat. 88-8824) and protocol. Please refer to Best Protocols: Protocol A: Two step protocol for (cytoplasmic) intracellular proteins located under the Resources Tab online. This can be used at 5 µL (0.125 µg) per test. A test is defined as the amount (µg) of antibody that will stain a cell sample in a final volume of 100 µL. Cell number should be determined empirically but can range from 10^5 to 10^8 cells/test. eFluor® 660 is a replacement for Alexa Fluor® 647. eFluor® 660 emits at 659 nm and is excited with the red laser (633 nm). Please make sure that your instrument is capable of detecting this fluorochome. Excitation: 633-647 nm; Emission: 668 nm; Laser: Red Laser. Filtration: 0.2 µm post-manufacturing filtered.

Reactivity

Human

Host

Mouse

Isotype

IgG

Antibody clone number

4B2G4

Vial size

25 Tests

Concentration

5 µL/Test

Storage

4° C, store in dark, DO NOT FREEZE!

References

Submitted references

Granzyme M targets host cell hnRNP K that is essential for human cytomegalovirus replication.

van Domselaar R, de Poot SA, Remmerswaal EB, Lai KW, ten Berge IJ, Bovenschen N
Cell death and differentiation 2013 Mar;20(3):419-29

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Human and mouse granzyme M display divergent and species-specific substrate specificities.

de Poot SA, Westgeest M, Hostetter DR, Van Damme P, Plasman K, Demeyer K, Broekhuizen R, Gevaert K, Craik CS, Bovenschen N
The Biochemical journal 2011 Aug 1;437(3):431-42

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A role for granzyme M in TLR4-driven inflammation and endotoxicosis.

Anthony DA, Andrews DM, Chow M, Watt SV, House C, Akira S, Bird PI, Trapani JA, Smyth MJ
Journal of immunology (Baltimore, Md. : 1950) 2010 Aug 1;185(3):1794-803

Flow cytometry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Normal human peripheral blood cells were stimulated for 3 days with Human IL-2 Recombinant Protein (Product # 14-8029-81), then cultured with Protein Transport Inhibitor Cocktail (Product # 00-4980-03) for an additional 5 hours. The cells were intracellularly stained with Anti-Human CD8 FITC (Product # 11-0087-42) and Mouse IgG1 Isotype Control eFluor® 660 (Product # 50-4714-82) (left) or Anti-Human Granzyme M eFluor® 660 (right) using the Intracellular Fixation and Permeabilization Buffer Set (Product # 88-8824-00) and protocol. Viable cells in the lymphocyte gate, as determined by Fixable Viability Dye eFluor® 450 (Product # 65-0863-14), were used for analysis.