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      FITC Rabbit Anti-Active Caspase-3
      FITC Rabbit Anti-Active Caspase-3
      Flow cytometric analysis of Active Caspase-3 expression by Apoptotic and Non-apoptotic Human Jurkat cells.  Cells from the Human Jurkat (T-cell leukemia, ATCC® TIB-152™) cell line were left untreated (Left histogram) or treated with 12 μM of Camptothecin for 6 hours to induce apoptosis (Right histogram). Cells were washed, fixed with BD Cytofix™ Fixation Buffer (Cat. No. 554655) and then permeabilized with BD Perm/Wash™ Buffer (Cat. No. 554723). The cells were subsequently stained in BD Perm/Wash™ Buffer with FITC Rabbit Anti-Active Caspase-3 antibody (Cat. No. 570334/570410) at 1 µg/test. The fluorescence histograms showing Active Caspase-3 expression were derived from gated events with the forward and side light-scatter characteristics of single cells. Flow cytometry and data analysis were performed using a BD LSRFortessa™ X-20 Cell Analyzer System and FlowJo™ Software. Data shown on this Technical Data Sheet are not lot specific.
      FITC Rabbit Anti-Active Caspase-3
      Flow cytometric analysis of Active Caspase-3 expression by Apoptotic and Non-apoptotic Mouse thymocytes.  C57BL/6 Mouse thymocytes were left untreated (Left histogram) or treated with 1 μM of Dexamethasone for 5 hours to induce apoptosis (Right histogram). Cells were washed, fixed with BD Cytofix™ Fixation Buffer (Cat. No. 554655) and then permeabilized with BD Perm/Wash™ Buffer (Cat. No. 554723). The cells were subsequently stained in BD Perm/Wash™ Buffer with FITC Rabbit Anti-Active Caspase-3 antibody (Cat. No. 570334/570410) at 1 µg/test. The fluorescence histograms showing Active Caspase-3 expression were derived from gated events with the forward and side light-scatter characteristics of single cells. Flow cytometry and data analysis were performed using a BD LSRFortessa™ X-20 Cell Analyzer System and FlowJo™ Software. Data shown on this Technical Data Sheet are not lot specific.
      FITC Rabbit Anti-Active Caspase-3 FITC Rabbit Anti-Active Caspase-3
      Flow cytometric analysis of Active Caspase-3 expression by Apoptotic and Non-apoptotic Human Jurkat cells.  Cells from the Human Jurkat (T-cell leukemia, ATCC® TIB-152™) cell line were left untreated (Left histogram) or treated with 12 μM of Camptothecin for 6 hours to induce apoptosis (Right histogram). Cells were washed, fixed with BD Cytofix™ Fixation Buffer (Cat. No. 554655) and then permeabilized with BD Perm/Wash™ Buffer (Cat. No. 554723). The cells were subsequently stained in BD Perm/Wash™ Buffer with FITC Rabbit Anti-Active Caspase-3 antibody (Cat. No. 570334/570410) at 1 µg/test. The fluorescence histograms showing Active Caspase-3 expression were derived from gated events with the forward and side light-scatter characteristics of single cells. Flow cytometry and data analysis were performed using a BD LSRFortessa™ X-20 Cell Analyzer System and FlowJo™ Software. Data shown on this Technical Data Sheet are not lot specific.
      Flow cytometric analysis of Active Caspase-3 expression by Apoptotic and Non-apoptotic Mouse thymocytes.  C57BL/6 Mouse thymocytes were left untreated (Left histogram) or treated with 1 μM of Dexamethasone for 5 hours to induce apoptosis (Right histogram). Cells were washed, fixed with BD Cytofix™ Fixation Buffer (Cat. No. 554655) and then permeabilized with BD Perm/Wash™ Buffer (Cat. No. 554723). The cells were subsequently stained in BD Perm/Wash™ Buffer with FITC Rabbit Anti-Active Caspase-3 antibody (Cat. No. 570334/570410) at 1 µg/test. The fluorescence histograms showing Active Caspase-3 expression were derived from gated events with the forward and side light-scatter characteristics of single cells. Flow cytometry and data analysis were performed using a BD LSRFortessa™ X-20 Cell Analyzer System and FlowJo™ Software. Data shown on this Technical Data Sheet are not lot specific.
      Product Details
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      BD Pharmingen™
      Apopain; CASP-3; CASP3; CC3; CPP-32; CPP32; Caspase 3; Yama
      Human (QC Testing), Mouse (Tested in Development)
      Rabbit IgG, κ
      Human Active Caspase-3 Fragment
      Intracellular staining (flow cytometry) (Routinely Tested)
      0.5 mg/ml
      836, 12367
      Aqueous buffered solution containing ≤0.09% sodium azide.
      RUO


      Preparation And Storage

      The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography. The antibody was conjugated to the dye under optimum conditions and unconjugated antibody and free dye were removed. Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze.
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      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 cell and BD® CompBeads to ensure that BD® CompBeads are appropriate for your specific cellular application.

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      Product Notices

      1. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
      2. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
      3. 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.
      4. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
      5. Species cross-reactivity detected in product development may not have been confirmed on every format and/or application.
      6. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
      7. For U.S. patents that may apply, see bd.com/patents.
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      570410 Rev. 1
      Antibody Details
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      C92-605.rMAb

      The C92-605.rMAb is a recombinant monoclonal antibody derived from C92-605 hybridoma cells. This antibody specifically recognizes 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 Caspase family of cysteine proteases play crucial roles 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, eg, D4-GDI and Bcl-2, and in the nucleus (eg, PARP).

      570410 Rev. 1
      Format Details
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      FITC
      Fluorescein (FITC) is part of the BD blue family of dyes. This is a small organic fluorochrome with an excitation maximum (Ex Max) at 494-nm and an emission maximum (Em Max) at 518-nm. FITC is designed to be excited by the Blue laser (488-nm) and detected using an optical filter centered near 520 nm (e.g., a 530/30-nm bandpass filter). Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
      altImg
      FITC
      Blue 488 nm
      494 nm
      518 nm
      570410 Rev.1
      Citations & References
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      Development References (5)

      1. 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
      2. 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
      3. 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: Intracellular Staining/Flow Cytometry). View Reference
      4. Winzler C, Fantinato M, Giordan M, Calore E, Basso G, Messina C. CD4(+) T regulatory cells are more resistant to DNA damage compared to CD4(+) T effector cells as revealed by flow cytometric analysis.. Cytometry A. 2011; 79(11):903-11. (Clone-specific: Intracellular Staining/Flow Cytometry). View Reference
      5. Zhu S, Zhang X, Weichert-Leahey N, et al. LMO1 Synergizes with MYCN to Promote Neuroblastoma Initiation and Metastasis.. Cancer Cell. 2017; 32(3):310-323.e5. (Clone-specific: Immunohistochemistry). View Reference
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      570410 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.

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