Invitrogen™ Granzyme B Monoclonal Antibody (NGZB), eFluor™ 660, eBioscience™, Invitrogen™

Catalog No.	Quantity
50-112-9101	100 μg

Catalog No. 50-112-9101 Supplier Invitrogen™ Supplier No. 50889882

Description

Rat Monoclonal Antibody

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 (cat. 88-8824) and protocol. Please refer to Best Protocols: Protocol A: Two step protocol for (cytoplasmic) intracellular proteins.

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.

Specifications

• Antigen: Granzyme B
• Applications: Flow Cytometry
• Classification: Monoclonal
• Clone: NGZB
• Concentration: 0.2 mg/mL
• Conjugate: eFluor 660
• Formulation: PBS with 0.09% sodium azide; pH 7.2
• Gene: GZMB
• Gene Accession No.: P04187
• Gene Alias: AI553453; C11; Cathepsin G-like 1; CCP1; CCP-1/C11; CCPI; CGL1; CGL-1; CSPB; CSP-B; Ctla1; Ctla-1; CTSGL1; cytotoxic cell protease 1; cytotoxic serine protease B; Cytotoxic T lymphocyte associated serine esterase 1; cytotoxic T-lymphocyte proteinase 2; cytotoxic T-lymphocyte-associated serine esterase 1; fragmentin; fragmentin 2; fragmentin-2; GLP I; GLP III; GLP-1; granzyme 2; granzyme B; granzyme B (granzyme 2, cytotoxic T-lymphocyte-associated serine esterase 1); granzyme B(G,H); Granzyme-2; GranzymeB; granzyme-like protein 1; granzyme-like protein I; GRB; GZB; Gzmb; HLP; human lymphocyte protein; Human lymphocyte protein (Hlp); Lymphocyte protease; Natural killer cell protease 1; OTTHUMP00000028189; RNKP-1; SECT; T-cell serine protease 1-3E
• Gene Symbols: GZMB
• Host Species: Rat
• Purification Method: Affinity chromatography
• Quantity: 100 μg
• Regulatory Status: RUO
• Primary or Secondary: Primary
• Gene ID (Entrez): 14939
• Target Species: Mouse
• Content And Storage: 4°C, store in dark, DO NOT FREEZE!
• Product Type: Antibody
• Form: Liquid
• Isotype: IgG2a κ

Frequently Asked Questions (FAQs)

I am unable to analyze my cells stained with eFluor Organic Dyes today. What options do I have?

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.

Can the eFluor Organic fluorochromes be used for intracellular staining?

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).

Is the eFluor 660 fluorochrome compatible with Anti-Cy5/Alexa Fluor 647 beads?

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.

What buffers are compatible with the eFluor Organic fluorochromes and eVolve QDots?

The eFluor Organic fluorochromes and eVolve QDots can be used with flow staining buffers containing PBS and protein.

What is the difference between eFluor Organic Dyes and eVolve Qdots?

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.

Are the eFluor Organic Dyes photo-labile?

As with other fluorochromes, we recommend minimal exposure to light to maintain optimal signal.

For Research Use Only.