Key Benefits

  • Simultaneous detection of mitochondrial membrane potential and caspase activity.
  • Readout – Flow cytometry, Fluorescent plate reader, Fluorescent microscopy .
  • Reliable: Yields both quantitative and qualitative results. Gives a strong positive signal.
  • The kit can be used in conjunction with other antibodies or stains.
  • Ease Of Use: No need to make cell lysates or run western blots.
  • Cell Permeable Reagents.

Additional information

Kit Size

25, 100


Poly Caspase, Caspase 3/7, Caspase 8, Caspase 9, Caspase 1

Caspase (poly, 3/7, 8, 9, 1) & Mitochondria Membrane Potential Detection – MitoCasp

Caspase enzymes specifically recognize a 4 amino acid sequence (on their substrate) which necessarily includes an aspartic acid residue. This residue is the target for the cleavage reaction, which occurs at the carbonyl end of the aspartic acid residue(6). Caspases can be detected via immunoprecipitation, immuno-blotting techniques using caspase specific antibodies, or by employing fluorogenic substrates which become fluorescent upon cleavage by the caspase. MitoCasp uses a novel approach to detect active caspases (7-9). The methodology is based on carboxyfluorescein (FAM) labeled fluoromethyl ketone (FMK)-peptide inhibitors of caspases. These inhibitors are cell permeable and non-cytotoxic. Once inside the cell, the inhibitor binds covalently to the active caspase (10). Cells that contain bound inhibitor can be analyzed by flow cytometry or fluorescence microscopy.

Cell Technology utilizes a cationic dye to visualize mitochondrial membrane potential (15-17). The cationic dye is cell permeable and has a strong fluorescent signal in the red region and exhibits low membrane potential independent (non specific) binding and toxicity. In healthy cells the cationic dye is accumulated by the mitochondria in proportion to the DeltaPsi (membrane potential). In most cell lines, accumulation of the cationic dye in the mitochondria results in a higher fluorescence intensity. In apoptotic cells, where the mitochondrial membrane potential is compromised, the cationic dye does not accumulated in the mitochondria and these cells exhibit a lower fluorescence signal. Utilizing these two reagents in combination Caspase activity and mitochondrial membrane potential can be analyzed simultaneously. Citations Identification of single-domain, Bax-spec

Jurkat cells were stimulated with Staurosporine for 3 hours (B) or DMSO (A). The cells were then stained with the MitoCasp kit according to the protocol. The cells were then washed twice and analyzed by flow cytometry: Ex:488nm Em: FL1 and FL2.

Fig A.
Healthy cells show a strong red fluorescence indicating intact mitochondria and no green fluorescence, indicating no active caspases.

Fig B.
Apoptotic cells show a loss of red fluorescence (y axis) indicating loss of mitochondrial membrane potential and positive green fluorescence (x axis) indicating active caspases.

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Selective cytotoxicity of Pancratistatin-related natural Amaryllidaceae alkaloids: evaluation of the activity of two new compounds, Sharda, Sood, Nair, et alCancer Cell IntV7 2007
OSU-03012, a Novel Celecoxib Derivative, Is Cytotoxic to Myeloma Cells and Acts through Multiple Mechanisms, Suvannasankha, Crean, et.alClinical Cancer Research13, pp 4750-4758, Aug 15, 2007
Galectin-3 but not galectin-1 induces mast cell death by oxidative stress and mitochondrial permeability transition Suzuki, Toshio Inoue, Tetsuro Yoshimaru and Chisei RaBiochimica et Biophysica Acta (BBA) - Molecular Cell ResearchVol 1783, issue 5, pp 924-934, 2008
Nitric oxide protects mast cells from activation-induced cell death: the role of the phosphatidylinositol-3 kinase-Akt-endothelial nitric oxide synthase pathway Inoue, Yoshihiro Suzuki, Tetsuro Yoshimaru and Chisei RaJournal of Leukocyte Biologyvol. 83 no. 5, pp1218-1229, 2008 doi:10.1189/jlb.1007667
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Wilson, K. P., J. F. Black, J. A. Thomson, E. E. Kim, J. P. Griffith, M. A. Navia, M. A. Murcko, S. P. Chambers, R. A. Aldape, S. A. Raybuck, and D. J. Livingston. 1994. Structure and mechanism of interleukin-1 beta converting enzyme. Nature 370: 270-275.
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Part # 4015Mitochondrial Memberane Potential Cationic Dye2-8C
Refer to Product DatasheetCaspase Detection Reagent (Poly Caspase, Caspase 3/7, Caspase 1, Caspase 8 or Caspase 9)2-8C
Part # 302810X Wash Buffer2-8C
Part # 30321X Dilution Buffer2-8C

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