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Invitrogen
Granzyme B Monoclonal Antibody (NGZB), PE-eFluor™ 610, eBioscience™
FIGURE: 1 / 37
Granzyme B Antibody (61-8898-82) in Flow
C57Bl/6 splenocytes were cultured with plate-bound Anti-Mouse CD3e Functional Grade Purified (Product # 16-0031-82) and Anti-Mouse CD28 Functional Grade Purified (Product # 16-0281-82) for 3 days, then cultured with Protein Transport Inhibitor Cocktail (Product # 00-4980-03) for an additional 5 hours. Cells were surface stained with Anti-Mouse CD8a FITC (Product # 11-0081-82) followed by intracellular staining with 0.125 µg of Rat IgG2a K Isotype Control PE-eFluor® 610 (Product # 61-4321-82) (left) or 0.125 µg of Anti-Mouse Granzyme B PE-e... View More
Product Details
61-8898-82
Applications
Tested Dilution
Publications
Product Specifications
Species Reactivity
Mouse
Published species
Mouse
Host/Isotype
Rat
/ IgG2a, kappa
Recommended Isotype Control
Class
Monoclonal
Type
Antibody
Clone
NGZB
Conjugate
Excitation/Emission Max
565/606 nm
View spectra
Form
Liquid
Concentration
0.2 mg/mL
Purification
Affinity chromatography
Storage buffer
PBS, pH 7.2
Contains
0.09% sodium azide
Storage conditions
4° C, store in dark, DO NOT FREEZE!
Shipping conditions
Ambient (domestic); Wet ice (international)
RRID
AB_2574670
Product Specific Information
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 followed by flow cytometric analysis.
Applications Tested: This NGZB antibody has been tested by intracellular staining and flow cytometric analysis of stimulated mouse splenocytes using the Intracellular Fixation & 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.
PE-eFluor® 610 can be excited with laser lines from 488-561 nm and emits at 607 nm. We recommend using a 610/20 band pass filter (equivalent to PE-Texas Red®). Please make sure that your instrument is capable of detecting this fluorochome.
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: 488-561 nm; Emission: 607 nm; Laser: Blue Laser, Green Laser, Yellow-Green Laser.
Filtration: 0.2 µm post-manufacturing filtered.
Target Information
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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Bioinformatics
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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