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Invitrogen™ Granzyme M Monoclonal Antibody (4B2G4), eFluor™ 660, eBioscience™
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 (Product # 88-8824-00) and protocol. Please refer to BestProtocols®: 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...
Specifications
Specifications
| Antigen | Granzyme M |
| Applications | Flow Cytometry |
| Classification | Monoclonal |
| Clone | 4B2G4 |
| Concentration | 5 μL/Test |
| Conjugate | eFluor 660 |
| Formulation | PBS with BSA and 0.09% sodium azide; pH 7.2 |
| Gene | GZMM |
| Gene Accession No. | P51124 |
| Gene Alias | granzyme M; granzyme M (lymphocyte met-ase 1); GZMM; Hu-Met-1; Lmet1; lymphoctye Met-ase 1; lymphocyte met-ase 1; MET1; Met-1 serine protease; Met-ase; MMET-1; natural killer cell granular protease; RNK-Met-1 |
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Frequently Asked Questions (FAQs)
Our options will depend on the samples you are analyzing.
If cell viability is not critical, you can store your stained samples at 4 degrees C or on ice overnight in the dark and analyze the following day.
For samples stained with eFluor organic fluorochromes, we recommend that cells be suspended in 100 uL of Flow Cytometry Staining Buffer (Cat. No. 00-4222) and 100 uL of eBioscience IC Fixation Buffer (Cat. No. 00-8222); samples can be incubated for up to 3 days at 4 degrees C in the dark. Alternatively, the 1-step Fix/Lyse Solution (Cat. No. 00-5333) can be used. This is a great option when working with whole blood but also works for other cell types.
Yes, the eFluor Organic fluorochromes can be used for intracellular staining. The eFluor organic fluorochromes maintain bright signal and require minimal changes in compensation when fixed with eBioscience IC Fixation Buffer (Cat. No. 00-8222-49) and Permeabilization Buffer (Cat. No. 00-8333-56) or 1-step Fix/Lyse Solution (Cat. No. 00-5333-54, 00-5333-57) (as compared to live cells).
Yes, in-house studies have demonstrated that the eFluor 660 fluorochrome is recognized by Anti-Cy5/Alexa Fluor 647 beads. Side by side studies with Alexa Fluor 647 versus eFluor 660 conjugated antibodies have demonstrated comparable results.
The eFluor Organic fluorochromes and eVolve QDots can be used with flow staining buffers containing PBS and protein.
The eFluor Organic Dyes (eFluor 450, APC-eFluor 780, PerCP-eFluor 710, eFluor 710) are conventional fluorochromes. In contrast, the eVolve line of products are Quantum dots.
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