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R718 Rabbit Anti-Active Caspase-3
R718 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 BD Horizon™ R718 Rabbit Anti-Active Caspase-3 antibody (Cat. No. 570305/570311) at 0.5 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.
R718 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 BD Horizon™ R718 Rabbit Anti-Active Caspase-3 antibody (Cat. No. 570305/570311) at 0.5 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 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 BD Horizon™ R718 Rabbit Anti-Active Caspase-3 antibody (Cat. No. 570305/570311) at 0.5 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 BD Horizon™ R718 Rabbit Anti-Active Caspase-3 antibody (Cat. No. 570305/570311) at 0.5 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 Horizon™
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)
0.2 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 unreacted dye was removed. Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze.

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

  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. Human donor specific background has been observed in relation to the presence of anti-polyethylene glycol (PEG) antibodies, developed as a result of certain vaccines containing PEG, including some COVID-19 vaccines. We recommend use of BD Horizon Brilliant™ Stain Buffer in your experiments to help mitigate potential background. For more information visit https://www.bdbiosciences.com/en-us/support/product-notices.
  6. Species cross-reactivity detected in product development may not have been confirmed on every format and/or application.
  7. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
  8. This product is provided under an Agreement between BIOTIUM and BD Biosciences. This product, and only in the amount purchased by buyer, may be used solely for buyer’s own internal research, in a manner consistent with the accompanying product literature. No other right to use, sell or otherwise transfer (a) this product, or (b) its components is hereby granted expressly, by implication or by estoppel. This product is for research use only. Diagnostic uses require a separate license from Biotium, Inc. For information on purchasing a license to this product including for purposes other than research, contact Biotium, Inc., 3159 Corporate Place, Hayward, CA 94545, Tel: (510) 265-1027. Fax: (510) 265-1352. Email: btinfo@biotium.com.
  9. Alexa Fluor™ is a trademark of Life Technologies Corporation.
  10. For U.S. patents that may apply, see bd.com/patents.
570305 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).

570305 Rev. 1
Format Details
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R718
The BD Horizon™ Red 718 (R718) Dye is part of the BD red family of dyes. It is a small organic fluorochrome with an excitation maximum (Ex Max) at 695-nm and an emission maximum (Em Max) at 718-nm. Driven by BD innovation, R718 is designed to be excited by the red laser (627–640-nm) and detected using an optical filter centered near 720-nm (e.g., a 720/40-nm bandpass filter). R718 is a brighter alternative to Alexa Fluor™ 700. R718 is also a bright small molecule alternative to APC-R700 with lower spread into the APC detector. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
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R718
Red 627-640 nm
695 nm
718 nm
570305 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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570305 Rev. 1

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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.