PE Rabbit Anti- Active Caspase-3

Consider alternate product [570183], clone C92-605.rMAb, which is manufactured with recombinant technology.

BD Pharmingen™ PE Rabbit Anti- Active Caspase-3

Name: PE Rabbit Anti- Active Caspase-3
Brand: BD Pharmingen™
SKU: 550821

Clone C92-605

(RUO)

Flow cytometric analysis of apoptotic and non-apoptotic populations for active caspase-3. Jurkat cells (Human T-cell leukemia; ATCC TIB-152) were left untreated (left panel) or treated with 4 µM of camptothecin for 4 hr to induce apoptosis (right panel). Cells were washed once in PBS, then fixed and permeabilized using the BD Cytofix/Cytoperm™ Kit (Cat. No. 554714) for 20 min at room temperature (RT), pelleted and washed with BD Perm/Wash™ buffer (component of Cat. No. 554714). Cells were subsequently stained with the PE rabbit anti- active caspase-3 antibody (clone C92-605). Cells were then washed and resuspended in BD Perm/Wash™ buffer before analyzing by flow cytometry. The results show that untreated cells were primarily negative for active caspase-3 (left panel, M1); whereas over one third of the treated cells were positive for active caspase-3 staining (right panel, M2).

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Product Details

Brand: BD Pharmingen™

Alternative Name: CPP32; Yama; Apopain

Reactivity: Human (QC Testing), Mouse (Tested in Development)

Isotype: Rabbit IgG

Immunogen: Human Active Caspase-3 Fragment

Application: Intracellular staining (flow cytometry) (Routinely Tested)

Target Molecular Weight: 17-22 kDa

Vol. Per Test: 20 µl

Storage Buffer: Aqueous buffered solution containing BSA and ≤0.09% sodium azide.

Regulatory Status: RUO

Preparation And Storage

The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography. The antibody was conjugated with R-PE under optimum conditions, and unconjugated antibody and free PE were removed. Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze.

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).
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.
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.

Companion Products

Fixation/Permeabilization Kit RUO

Size 250 Tests Cat No. 554714

550821 Rev. 3

Antibody Details

C92-605

The caspase family of cysteine proteases plays a key role in apoptosis and inflammation. Caspase-3 is a key protease that is activated during the early stages of apoptosis and, like other members of the caspase family, is synthesized as an inactive pro-enzyme that is processed in cells undergoing apoptosis by self-proteolysis and/or cleavage by another protease. The processed forms of caspases consist of large (17-22 kDa) and small (10-12 kDa) subunits which associate to form an active enzyme. Active caspase-3, a marker for cells undergoing apoptosis, consists of a heterodimer of 17 and 12 kDa subunits which is derived from the 32 kDa pro-enzyme. Active caspase-3 proteolytically cleaves and activates other caspases, as well as relevant targets in the cytoplasm, e.g., D4-GDI and Bcl-2, and in the nucleus (e.g. PARP). This antibody has been reported to specifically recognize the active form of caspase-3 in human and mouse cells. It has not been reported to recognize the pro-enzyme form of caspase-3.

550821 Rev. 3

Format Details

R-Phycoerythrin (PE), is part of the BD family of Phycobiliprotein dyes. This fluorochrome is a multimeric fluorescent phycobiliprotein with excitation maximum (Ex Max) of 496 nm and 566 nm and an emission maximum (Em Max) at 576 nm. PE is designed to be excited by the Blue (488 nm), Green (532 nm) and Yellow-Green (561 nm) lasers and detected using an optical filter centered near 575 nm (e.g., a 575/26-nm bandpass filter). As PE is excited by multiple lasers, this can result in cross-laser excitation and fluorescence spillover on instruments with various combinations of Blue, Green, and Yellow-Green lasers. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.

Format: PE

Excitation Source: Yellow-Green 488 nm, 532 nm, 561 nm

Excitation Max: 496 nm, 566 nm

Emission Max: 576 nm

550821 Rev.3

Citations & References

View product citations for antibody "550821" on CiteAb

Development References (3)

Dai C, Krantz SB. Interferon gamma induces upregulation and activation of caspases 1, 3, and 8 to produce apoptosis in human erythroid progenitor cells. Blood. 1999; 93(10):3309-3316. (Biology). View Reference
Fujita N, Tsuruo T. Involvement of Bcl-2 cleavage in the acceleration of VP-16-induced U937 cell apoptosis. Biochem Biophys Res Commun. 1998; 246(2):484-488. (Biology). View Reference
Thornberry NA, Lazebnik Y. Caspases: enemies within. Science. 1998; 281(5381):1312-1316. (Biology). View Reference

550821 Rev. 3

Please refer to Support Documents for Quality Certificates

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.