BD Pharmingen™ PE Mouse Anti-Human Granzyme A Set
Flow cytometric analysis of Granzyme A expression in human peripheral blood lymphocytes. Human peripheral blood mononuclear cells (PBMC) were fixed using BD Cytofix/Cytoperm™ Fixation and Permeabilization Solution (Cat. No. 554722). The cells were then washed and stained in BD Perm/Wash™ Buffer (Cat. No. 554723) with either PE Mouse IgG1 Isotype Control (Component No. 51-13855X-6; dashed line histogram) or PE Mouse Anti-Human Granzyme A antibody (Component No. 51-68395X; solid line histogram). The fluorescence histogram showing the expression of Granzyme A (or Ig Isotype control staining) was derived from gated events with the forward and side light-scatter characteristics of intact lymphocytes.
Flow cytometric analysis of Granzyme A expression in human peripheral blood lymphocytes. Human peripheral blood mononuclear cells (PBMC) were fixed using BD Cytofix/Cytoperm™ Fixation and Permeabilization Solution (Cat. No. 554722). The cells were then washed and stained in BD Perm/Wash™ Buffer (Cat. No. 554723) with either PE Mouse IgG1 Isotype Control (Component No. 51-13855X-6; dashed line histogram) or PE Mouse Anti-Human Granzyme A antibody (Component No. 51-68395X; solid line histogram). The fluorescence histogram showing the expression of Granzyme A (or Ig Isotype control staining) was derived from gated events with the forward and side light-scatter characteristics of intact lymphocytes.
Description
The primary mechanism by which cytotoxic T cells eliminate virally infected cells is by granule exocytosis. The release of cytotoxic granule contents by cytotoxic T lymphocytes (CTL) triggers apoptotic target cell death. CTL granules contain a pore-forming protein, perforin, and a group of serine proteases called granzymes. In the classic model, perforins create holes in the target cell membrane, allowing entrance of the granzymes. Granzyme A and B are the predominant granzymes activated after CTL activation, but each act via an independent apoptotic pathway; granzyme B is activated immediately, while granzyme A acts hours later. Granzyme B does not induce cleavage of caspase-3, lamin B, rho-GTPase or PARP, but does cleave DNA-PKcs and nuclear mitotic apparatus protein (NuMA). Studies involving mice which are deficient in both granzyme A and B suggest a model whereby the granzyme B pathway may have evolved as the major apoptotic pathway with the granzyme A pathway acting as a backup. However, further research is needed to delineate the components of the distinct pathways.
Preparation And Storage
Recommended Assay Procedures
BD™ CompBeads can be used as surrogates to assess fluorescence spillover (Compensation). When fluorochrome conjugated antibodies are bound to BD™ CompBeads, they have spectral properties very similar to cells. However, for some fluorochromes there can be small differences in spectral emissions compared to cells, resulting in spillover values that differ when compared to biological controls. It is strongly recommended that when using a reagent for the first time, users compare the spillover on cells and BD CompBeads to ensure that BD™ CompBeads are appropriate for your specific cellular application.
Product Notices
- This reagent has been pre-diluted for use at the recommended Volume per Test. We typically use 1 × 10^6 cells in a 100-µl experimental sample (a test).
- An isotype control should be used at the same concentration as the antibody of interest.
- Caution: Sodium azide yields highly toxic hydrazoic acid under acidic conditions. Dilute azide compounds in running water before discarding to avoid accumulation of potentially explosive deposits in plumbing.
- Source of all serum proteins is from USDA inspected abattoirs located in the United States.
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
| Description | Quantity/Size | Part Number | EntrezGene ID |
|---|---|---|---|
| N/A | 100.0 | N/A | N/A |
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Global - Refer to manufacturer's instructions for use and related User Manuals and Technical data sheets before using this products as described
Comparisons, where applicable, are made against older BD Technology, manual methods or are general performance claims. Comparisons are not made against non-BD technologies, unless otherwise noted.
For Research Use Only. Not for use in diagnostic or therapeutic procedures.
Refer to manufacturer's instructions for use and related User Manuals and Technical Data Sheets before using this product as described.
Comparisons, where applicable, are made against older BD technology, manual methods or are general performance claims. Comparisons are not made against non-BD technologies, unless otherwise noted.