Granzyme B Antibody, anti-human/mouse/rat, REAfinity™

Name: Granzyme B Antibody, anti-human/mouse/rat, REAfinity™
Availability: InStock

Granzyme B Antibody, anti-human/mouse/rat, REAfinity™Extended ValidationRecombinant

Clone:

REA226

Type of antibody:

Primary antibodies, Recombinant antibodies

Isotype:

recombinant human IgG1

Applications:

ICFC, MC

Alternative names:

GZMB, CCPI, CGL-1, CSP-B, CTLA-1, CTSGL1, HLP, SECT, Granzyme 2

Intracellular flow cytometry applications forGranzyme B Antibody, anti-human/mouse/rat, REAfinity™

APC
FITC
PE
Vio B515

Conjugate:

APC

Recommended dilution:

1:50

Remark:

The antibody is suited for staining of formaldehyde-fixed cells.

Tested:

QC tested

Products used:

130-120-773, 130-120-703

Protocols:

Intracellular flow cytometry staining protocol (Inside Stain Kit 1:50)

Description:

Human peripheral blood mononuclear cells (PBMCs) were fixed, permeabilized and stained with Anti-Granzyme A antibodies and CD8 antibodies. Cells were then analyzed by flow cytometry using the MACSQuant^®Analyzer. The specificity of the conjugated antibodies was confirmed by blocking the binding to the ligand,using pure unconjugated antibodies (left). Cell debris were excluded from the analysis based on scatter signals.

Mass cytometry applications forGranzyme B Antibody, anti-human/mouse/rat, REAfinity™

pure

Conjugate:

pure

Reported:

32676612,

Products used:

130-108-055

Extended validation for Granzyme B Antibody, anti-human/mouse/rat, REAfinity™

Specificity

Epitope competition

In order to compare the epitope specificity of an antibody, the clone being used is compared with other known clones recognizing the same antigen in a competition assay.

Other clones	Overlap in epitope recognition with REA226
GB11	++
QA16A02	+
Cells were incubated with an excess of purified unconjugated Granzyme B (REA226) antibody followed by staining with fluorochrome-conjugated antibodies of other known clones against the same marker. Based on the fluorescence signal obtained, the clones were identified as recognizing completely overlapping (++), partially overlapping (+), or completely different epitopes (-) of the marker.

Sensitivity

Performance comparison

Selected fluorochrome conjugated antibodies from Miltenyi Biotec were compared to commercially available hybridoma clones in flow cytometry analysis.

Flow cytometric comparison of different clones for Granzyme B. Human peripheral blood mononuclear cells (PBMCs) were stained with a suitable counterstaining. Cells were then fixed and permeabilized followed by a staining with Granzyme B. As a control, Granzyme B antibody staining was omitted and cells were measured in the same channels. Flow cytometry was performed with the MACSQuant^® Analyzer. Cell debris and cell doublets were excluded from the analysis based on scatter signals. No FcR Blocking Reagent was used. The recommended titers of respective antibodies from different suppliers were used.

Flow cytometric comparison of different clones for Granzyme B. Human peripheral blood mononuclear cells (PBMCs) were stained with a suitable counterstaining. Cells were then fixed and permeabilized followed by a staining with Granzyme B. As a control, Granzyme B antibody staining was omitted and cells were measured in the same channels. Flow cytometry was performed with the MACSQuant^® Analyzer. Cell debris and cell doublets were excluded from the analysis based on scatter signals. No FcR Blocking Reagent was used. The recommended titers of respective antibodies from different suppliers were used.

Flow cytometric comparison of different clones for Granzyme B. Human peripheral blood mononuclear cells (PBMCs) were stained with a suitable counterstaining. Cells were then fixed and permeabilized followed by a staining with Granzyme B. As a control, Granzyme B antibody staining was omitted and cells were measured in the same channels. Flow cytometry was performed with the MACSQuant^® Analyzer. Cell debris and cell doublets were excluded from the analysis based on scatter signals. No FcR Blocking Reagent was used. The recommended titers of respective antibodies from different suppliers were used.

Flow cytometric comparison of different clones for Granzyme B. Human peripheral blood mononuclear cells (PBMCs) were stained with a suitable counterstaining. Cells were then fixed and permeabilized followed by a staining with Granzyme B. As a control, Granzyme B antibody staining was omitted and cells were measured in the same channels. Flow cytometry was performed with the MACSQuant^® Analyzer. Cell debris and cell doublets were excluded from the analysis based on scatter signals. No FcR Blocking Reagent was used. The recommended titers of respective antibodies from different suppliers were used.

Specifications for Granzyme B Antibody, anti-human/mouse/rat, REAfinity™

Overview

Clone REA226 recognizes granzyme B, a 32 kDa member of serine protease family, which is involved in cell lysis mediated by cytotoxic T lymphocytes (CTLs) and NK cells. Also known as granule enzymes, granzymes comprise majority of the mass of cytolytic granules in cytotoxic T lymphocytes (CTLs) and NK cells. Granzyme B is one of the five granzymes expressed by humans. NK cells and γδ T cells express granzymes constitutively, whereas expression in T lymphocytes is stimulation dependent. Main function attributed to granzymes is to indue apoptotic death of infected and transformed cells, where granzyme B enters into the target cell in a perforin dependent manner and via cation-independent mannose-6-phosphate receptor (CI-MPR). Granzyme B cleaves cellular substrates such as procaspase-3 and Bid, which then initiate pathways involved in apoptotic cell death. Granzyme B is suggested to be the only mammalian serine protease that prefers acidic side chains.

Additional information: Clone REA226 displays negligible binding to Fc receptors.

Alternative names

GZMB, CCPI, CGL-1, CSP-B, CTLA-1, CTSGL1, HLP, SECT, Granzyme 2

Detailed product information

Technical specifications

Clone	REA226
Clonality	monoclonal
Isotype	recombinant human IgG1
Isotype control	REA Control Antibody (I), human IgG1
Host	human cell line
Type of antibody	Primary antibodies, Recombinant antibodies
Species	human, mouse, rat
Antigen	Granzyme B
Alternative names of antigen	GZMB, CCPI, CGL-1, CSP-B, CTLA-1, CTSGL1, HLP, SECT, Granzyme 2
Distribution of antigen	T cells, NK cells
Entrez Gene ID	3002, 14939, 171528
RRID	AB_2733664, AB_2727639, AB_2727564, AB_2784341, AB_2784340, AB_2659980, AB_2922317, AB_2922311, AB_2733663

Resources for Granzyme B Antibody, anti-human/mouse/rat, REAfinity™

Certificates

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References for Granzyme B Antibody, anti-human/mouse/rat, REAfinity™

Publications

Grossman, W. J. et al. (2004) Differential expression of granzymes A and B in human cytotoxic lymphocyte subsets and T regulatory cells. Blood 104: 2840-2848
Müllbacher, A. et al. (1999) Granzymes are the essential downstream effector molecules for the control of primary virus infections by cytolytic leukocytes. Proc. Natl. Acad. Sci. U.S.A. 96(13): 13950 -13955
Trapani, J. A. (2001) Granzymes: a family of lymphocyte granule serine proteases. Genome Biol. 2(12): 3014.1-3014.7

Applications

Extended validation