BV650 Rabbit Anti- Active Caspase-3

Consider alternate product [570179], clone C92-605.rMAb, which is manufactured with recombinant technology.

BD Horizon™ BV650 Rabbit Anti- Active Caspase-3

Name: BV650 Rabbit Anti- Active Caspase-3
Brand: BD Horizon™
SKU: 564096

Clone C92-605

(RUO)

Flow cytometric analysis of Active Caspase-3 expressed by apoptotic Jurkat cells. Cells from the human Jurkat (Acute T cell leukemia, ATCC TIB-152) cell line were untreated (dashed line histogram) or treated (solid line histogram) with 4 μM camptothecin for 4 hr to induce apoptosis. The cells were washed once in Dulbecco's PBS, fixed and permeabilized using the BD Cytofix/Cytoperm™ Fixation/Permeabilization Solution Kit (Cat. No. 554714; 20 min at room RT), pelleted and washed with BD Perm/Wash™ Buffer (a component of the kit). The cells were subsequently stained with the BD Horizon™ BV650 Rabbit Anti-Active Caspase-3 antibody (Cat. No. 564096). The cells were then washed, resuspended in BD Perm/Wash™ Buffer, and analyzed using a BD™ LSR II Flow Cytometer System. The fluorescence histograms were derived from gated events with the forward and side light-scatter characteristics of intact Jurkat cells.

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Product Details

Brand: BD Horizon™

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)

Vol. Per Test: 5 µl

RRID: AB_2738589

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

Regulatory Status: RUO

Preparation And Storage

Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze. The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography. The antibody was conjugated with BD Horizon™ BV650 under optimum conditions, and unconjugated antibody and free BD Horizon™ BV650 were removed.

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).
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.
Species cross-reactivity detected in product development may not have been confirmed on every format and/or application.
Alexa Fluor® is a registered trademark of Molecular Probes, Inc., Eugene, OR.
Brilliant Violet™ 650 is a trademark of Sirigen.
For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.

Companion Products

Fixation/Permeabilization Kit RUO

Size 250 Tests Cat No. 554714

564096 Rev. 1

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.

The antibody was conjugated to BD Horizon™ BV650 which is part of the BD Horizon Brilliant™ Violet family of dyes. This dye is a tandem fluorochrome of BD Horizon BV421 with an Ex Max of 405-nm and an acceptor dye with an Em Max at 650-nm. BD Horizon BV650 can be excited by the violet laser and detected in a filter used to detect APC-like dyes (eg, 660/20-nm filter). Due to the excitation and emission characteristics of the acceptor dye, there will be spillover into the APC and Alexa Fluor® 700 detectors. However, the spillover can be corrected through compensation as with any other dye combination.

564096 Rev. 1

Format Details

BV650

The BD Horizon Brilliant Violet™ 650 (BV650) Dye is part of the BD Horizon Brilliant Violet™ family of dyes. This tandem fluorochrome is comprised of a BV421 donor with an excitation maximum (Ex Max) of 406-nm and an acceptor dye with an emission maximum (Em Max) at 649-nm. BV650, driven by BD innovation, is designed to be excited by the violet laser (405-nm) and detected using an optical filter centered near 650-nm (e.g., a 660/20-nm bandpass filter). The acceptor dye can be excited by the Red (628–640-nm) laser resulting in cross-laser excitation and fluorescence spillover. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.

Format: BV650

Excitation Source: Violet 405 nm

Excitation Max: 406 nm

Emission Max: 649 nm

564096 Rev.1

Citations & References

Development References (6)

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
Dukers DF, Oudejans JJ, Vos W, ten Berge RL, Meijer CJ. Apoptosis in B-cell lymphomas and reactive lymphoid tissues always involves activation of caspase 3 as determined by a new in situ detection method. J Pathol. 2002; 196(3):307-315. (Clone-specific: Immunohistochemistry, Immunoprecipitation). 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
Ohsawa S, Hamada S, Yoshida H, Miura M. Caspase-mediated changes in histone H1 in early apoptosis: prolonged caspase activation in developing olfactory sensory neurons. Cell Death Differ. 2008; 15(9):1429-1439. (Clone-specific: Fluorescence microscopy, Immunofluorescence, Western blot). View Reference
Pettersen RD, Bernard G, Olafsen MK, Pourtein M, Lie SO. CD99 signals caspase-independent T cell death. J Immunol. 2001; 166(8):4931-4942. (Clone-specific: Flow cytometry). View Reference
Thornberry NA, Lazebnik Y. Caspases: enemies within. Science. 1998; 281(5381):1312-1316. (Biology). View Reference

View All (6)

564096 Rev. 1

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.