CaspSCREENTM Flow Cytometric Apoptosis Detection Kit

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Antigen
  • CG5370
  • DCP-1
  • DCP1
  • Dcp1
  • Dmel\\CG5370
  • ccp1
  • dcp-1
  • dcp1
  • l(2)01862
  • l(2)02132
  • Death caspase-1
  • Dcp-1
Reactivity
Mammalian
2
1
Application
Apoptosis Detection (AD), Flow Cytometry (FACS)
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Purpose The CaspSCREENTM Flow Cytometric Apoptosis Detection Kit provides a convenient means for detecting activation of caspases by flow cytometry in living cells. The assay is based on the cleavage of (aspartyl)2-Rhodamine 110 (D₂R), a reported substrate for members of caspase family proteases. The caspase substrate D₂R is non-fluorescent, however, upon cleavage of the substrate by cellular caspases, the released rhodamine 110 gives rise to fluorescence that can be measured at excitation of 488 nm and emission of 530 nm.
Brand CaspSCREEN™
Sample Type Cell Samples
Detection Method Fluorometric
Specificity The CaspSCREEN TM Flow Cytometric Apoptosis Detection Kit provides a convenient means for detecting activation of caspases by flow cytometry in living cells. The assay is based on the cleavage of (aspartyl) 2 -Rhodamine 110 (D 2 R), a reported substrate for members of caspase family proteases. The caspase substrate D 2 R is non-fluorescent, however, upon cleavage of the substrate by cellular caspases, the released rhodamine 110 gives rise to fluorescence that can be measured at excitation of 488 nm and emission of 530 nm. As the D 2 R is more cell-permeable than other fluorometric caspase substrates, activation of caspases can easily be measured in intact cells by flow cytometry.
Characteristics CaspSCREENTM Flow Cytometric Apoptosis Detection Kit: Simple & Convenient Kit for Detecting Activation of Caspases by Flow Cytometry in Living Cells.
Components D₂R Reagent
DTT (1 M)
D₂R Incubation Buffer
Alternative Name Caspase (CASP ELISA Kit Abstract)
Background Activation of members of caspase-family proteases plays a key role in apoptosis. The CaspSCREENTM Flow Cytometric Apoptosis Detection Kit provides a convenient means for detecting activation of caspases by flow cytometry in living cells. The assay is based on the cleavage of (aspartyl)2-Rhodamine 110 (D₂R), a reported substrate for members of caspase family proteases. The caspase substrate D₂R is non-fluorescent, however, upon cleavage of the substrate by cellular caspases, the released rhodamine 110 gives rise to fluorescence that can be measured at excitation of 488 nm and emission of 530 nm.
Application Notes Detection of activated caspases in living cells.
Comment

Further details regarding sample type: Live cells

Protocol 1. Induce apoptosis in cells by desired method. Concurrently incubate a control culture without induction.
2. Count cells and pellet 1 x 10 5 cells.
3. Resuspend cells in 0.3 mL of D 2 R Incubation Buffer.
3. Add 3 µL of the 1 M DTT (10 mM final concentration).
4. Add 1 µL of the D 2 R Reagent.
5. Incubate at 37 °C for 10-20 min in the dark.
6. Analyzing cells by flow cytometry using FL-1 channel (Ex/Em = 488/530 nm).
Restrictions For Research Use only
Storage -20 °C
Storage Comment Store kit at -20 °C (Store the Incubation Buffer at 4 °C after opening).
Expiry Date 6-12 months
Product cited in: Jin, Strich, Cooper: "Slt2p phosphorylation induces cyclin C nuclear-to-cytoplasmic translocation in response to oxidative stress." in: Molecular biology of the cell, Vol. 25, Issue 8, pp. 1396-407, 2014 (PubMed).

Hao, Cheng, Clancy, Nguyen: "Caspofungin kills Candida albicans by causing both cellular apoptosis and necrosis." in: Antimicrobial agents and chemotherapy, Vol. 57, Issue 1, pp. 326-32, 2012 (PubMed).

Katoh, Osanai, Tomita, Okumura: "Brain natriuretic peptide is released from human astrocytoma cell line U373MG under hypoxia: a possible role in anti-apoptosis." in: The Journal of endocrinology, Vol. 208, Issue 1, pp. 51-7, 2010 (PubMed).

Váchová, Palková: "Physiological regulation of yeast cell death in multicellular colonies is triggered by ammonia." in: The Journal of cell biology, Vol. 169, Issue 5, pp. 711-7, 2005 (PubMed).