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Description: This NGZB monoclonal antibody reacts with mouse Granzyme B, which is a member of the granzyme serine protease family. Granzyme B 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, and 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. Granzyme B 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. This NGZB monoclonal antibody does not crossreact to human Granzyme B nor is staining blocked with GB11, suggesting it recognizes a different epitope.
Applications Reported: This NGZB antibody has been reported for use in intracellular staining and flow cytometric analysis.
Applications Tested: This NGZB antibody has been tested by intracellular staining and flow cytometric analysis of mouse splenocytes using the Intracellular Fixation and Permeabilization Buffer Set (Product # 88-8824-00) and protocol. Please refer to Best Protocols: Protocol A: Two step protocol for (cytoplasmic) intracellular proteins. This can be used at less than or equal to 0.25 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. It is recommended that the antibody be carefully titrated for optimal performance in the assay of interest.
APC-eFluor™ 780 emits at 780 nm and is excited with the Red laser (633 nm). Please make sure that your instrument is capable of detecting this fluorochrome.
Light sensitivity: This tandem dye is sensitive to photo-induced oxidation. Please protect this vial and stained samples from light.
Fixation: Samples can be stored in IC Fixation Buffer (Product # 00-8222) (100 µL of cell sample + 100 µL of IC Fixation Buffer) or 1-step Fix/Lyse Solution (Product # 00-5333) for up to 3 days in the dark at 4°C with minimal impact on brightness and FRET efficiency/compensation. Some generalizations regarding fluorophore performance after fixation can be made, but clone specific performance should be determined empirically.
Excitation: 633-647 nm; Emission: 780 nm; Laser: Red Laser.
Granzyme B is a member of the granzyme serine protease family, and is found in the granules of cytotoxic T cells and NK cells. Granzyme B has been described as CGL1 (cathepsin G-like-1), a serine protease expressed only in cytotoxic T-lymphocytes after cell activation, and 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. Granzyme B is crucial for the rapid induction of target cell death by apoptosis, induced by interaction with cytotoxic T cells. The receptor involved in this process has been identified as mannose 6-phosphate receptor which functions as a death receptor for Granzyme B during cytotoxic T cell-induced apoptosis. Granzyme B enters target cells to cleave caspase-3 and initiate the caspase cascade leading to DNA fragmentation and apoptosis. Granzyme B can also act through a mitochondrial apoptosis pathway by cleaving the Bid protein. Granzymes are neutral serine proteases, which are stored in specialized lytic granules of cytotoxic T lymphocytes (CTLs) and in natural killer (NK) cells. A number of granzymes (A to G) have been isolated and cloned from mouse CTLs and NK cells, however in man, fewer have been cloned and identified.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
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Protein Aliases: CCP1; CTLA-1; Cytotoxic cell protease 1; Cytotoxic T lymphocyte associated serine esterase 1; Fragmentin-2; Granzyme B(G,H); GranzymeB; Human lymphocyte protein (Hlp); OTTHUMP00000028189
Gene Aliases: AI553453; CCP-1/C11; CCP1; Ctla-1; Ctla1; GZB; Gzmb
UniProt ID: (Mouse) P04187
Entrez Gene ID: (Mouse) 14939
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