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      BV605 Rabbit Anti- Active Caspase-3
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      Consider alternate product [570538], clone C92-605.rMAb, which is manufactured with recombinant technology.
      BV605 Rabbit Anti- Active Caspase-3
      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™ BV605 Rabbit Anti-Active Caspase-3 antibody (Cat. No. 564094). 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.
      BV605 Rabbit Anti- Active Caspase-3
      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™ BV605 Rabbit Anti-Active Caspase-3 antibody (Cat. No. 564094). 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.
      Product Details
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      BD Horizon™
      CPP32; Yama; Apopain
      Human (QC Testing), Mouse (Tested in Development)
      Rabbit IgG
      Human Active Caspase-3 Fragment
      Intracellular staining (flow cytometry) (Routinely Tested)
      5 µl
      AB_2738587
      Aqueous buffered solution containing BSA and ≤0.09% sodium azide.
      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™ BV605 under optimum conditions, and unconjugated antibody and free BD Horizon™ BV605 were removed.
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      Product Notices

      1. 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).
      2. 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.
      3. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
      4. Please observe the following precautions: Absorption of visible light can significantly alter the energy transfer occurring in any tandem fluorochrome conjugate; therefore, we recommend that special precautions be taken (such as wrapping vials, tubes, or racks in aluminum foil) to prevent exposure of conjugated reagents, including cells stained with those reagents, to room illumination.
      5. Although every effort is made to minimize the lot-to-lot variation in the efficiency of the fluorochrome energy transfer, differences in the residual emission from BD Horizon™ BV421 may be observed. Therefore, we recommend that individual compensation controls be performed for every BD Horizon™ BV605 conjugate.
      6. Species cross-reactivity detected in product development may not have been confirmed on every format and/or application.
      7. CF™ is a trademark of Biotium, Inc.
      8. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
      9. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
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      564094 Rev. 2
      Antibody Details
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      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.

      This antibody is conjugated to BD Horizon BV605 which is part of the BD Horizon Brilliant™ Violet family of dyes. With an Ex Max of 407-nm and Em Max of 602-nm, BD Horizon BV605 can be excited by a violet laser and detected with a standard 610/20-nm filter set. BD Horizon BV605 is a tandem fluorochrome of BD Horizon BV421 and an acceptor dye with an Em max at 605-nm. Due to the excitation of the acceptor dye by the green (532 nm) and yellow-green (561 nm) lasers, there will be significant spillover into the PE and BD Horizon PE-CF594 detectors off the green or yellow-green lasers. BD Horizon BV605 conjugates are very bright, often exhibiting brightness equivalent to PE conjugates and can be used as a third color off of the violet laser.

      For optimal and reproducible results, BD Horizon Brilliant Stain Buffer should be used anytime two or more BD Horizon Brilliant dyes are used in the same experiment.  Fluorescent dye interactions may cause staining artifacts which may affect data interpretation.  The BD Horizon Brilliant Stain Buffer was designed to minimize these interactions.  More information can be found in the Technical Data Sheet of the BD Horizon Brilliant Stain Buffer (Cat. No. 563794).

      564094 Rev. 2
      Format Details
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      BV605
      The BD Horizon Brilliant Violet™ 605 (BV605) 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 407-nm and an acceptor dye with an emission maximum (Em Max) at 605-nm. BV605, driven by BD innovation, is designed to be excited by the violet laser (405-nm) and detected using an optical filter centered near 610-nm (e.g., a 610/20-nm bandpass filter). The acceptor dye can be excited by the yellow-green (561-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.
      altImg
      BV605
      Violet 405 nm
      407 nm
      605 nm
      564094 Rev.2
      Citations & References
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      Development References (6)

      1. 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
      2. 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
      3. 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
      4. 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
      5. 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
      6. Thornberry NA, Lazebnik Y. Caspases: enemies within. Science. 1998; 281(5381):1312-1316. (Biology). View Reference
      View All (6) View Less
      564094 Rev. 2

      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.

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