CaspGLOWTM Red Active Caspase-8 Staining Kit

Details for Product No. ABIN412033, Supplier: Log in to see
Antigen
  • casp8-A
  • xCaspase-8
  • casp8
  • CASP8
  • CASP-8
  • FLICE
  • MACH
  • Mch5
  • ALPS2B
  • CAP4
  • Casp-8
  • MCH5
  • caspase-8
  • zgc:92075
  • caspase 8, apoptosis-related cysteine peptidase
  • death related ced-3/Nedd2-like protein
  • caspase-8
  • caspase 8
  • casp8
  • Dredd
  • CASP8
  • AaeL_AAEL014148
  • LOC100222284
  • Casp8
  • LOC100348477
Reactivity
Mammalian
26
15
15
4
4
2
1
1
1
1
1
1
1
Application
Detection (D), Fluorescence Microscopy (FM), Flow Cytometry (FACS)
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Purpose The CaspGLOWTM Red Active Caspase-8 Staining Kit provides a convenient means for detecting activated caspase-8 in living cells. The assay utilizes a caspase-8 inhibitor IETD-FMK conjugated to sulfo-rhodamine (Red-IETD-FMK) as the fluorescent marker. Red-IETD-FMK is cell permeable, nontoxic, and irreversibly binds to activated caspase-8 in apoptotic cells.
Brand CaspGLOW™
Sample Type Cell Samples
Detection Method Fluorometric
Specificity The CaspGLOW™ Red Active Caspase-8 Staining Kit provides a convenient means for detecting activated caspase-8 in living cells. The assay utilizes a caspase-8 inhibitor IETD-FMK conjugated to sulfo-rhodamine (Red-IETD-FMK) as the fluorescent marker. Red-IETD-FMK is cell permeable, nontoxic, and irreversibly binds to activated caspase-8 in apoptotic cells. The fluorescence label allows detection of activated caspase-8 in apoptotic cells directly by fluorescence microscopy, flow cytometry, or fluorescence plate reader.
Characteristics CaspGLOWTM Red Active Caspase-8 Staining Kit: Convenient & Sensitive Kit to Detect Activated Caspase-8 in Living Cells. Detection Method: Fluorescence Microscopy, Flow Cytometry or Fluorescence Plate Reader.
Components Red-IETD-FMK
Wash Buffer
Z-VAD-FMK
Alternative Name Caspase-8 (CASP8 ELISA Kit Abstract)
Background Activation of caspases plays a central role in apoptosis. The CaspGLOWTM Red Active Caspase-8 Staining Kit provides a convenient means for detecting activated caspase-8 in living cells. The assay utilizes a caspase-8 inhibitor IETD-FMK conjugated to sulfo-rhodamine (Red-IETD-FMK) as the fluorescent marker. Red-IETD-FMK is cell permeable, nontoxic, and irreversibly binds to activated caspase-8 in apoptotic cells.
Research Area Apoptosis/Necrosis
Application Notes Detection of activated caspase-8 in living cells.
Comment

Further details regarding sample type: Live cells

Protocol A. Staining Procedure:
1. Induce apoptosis in cells (1 x 10^6 /mL) by desired method. Concurrently incubate a control culture without induction. An additional negative control can be prepared by adding the caspase inhibitor Z-VAD-FMK at 1 µL/mL to an induced culture to inhibit caspase activation.
2. Aliquot 300 µL each of the induced and control cultures into eppendorf tubes.
3. Add 1 µL of Red-IETD-FMK into each tube and incubate for 0.5-1 hour at 37 °C incubator with 5 % CO 2.
4. Centrifuge cells at 3000 rpm for 5 minutes and remove supernatant.
5. Resuspend cells in 0.5 mL of Wash Buffer, and centrifuge again.
6. Repeat Step
5. Proceed to B, C, or D depending on methods of analysis. B. Quantification by Flow Cytometry: For flow cytometric analysis, resuspend cells in 300 µL of Wash buffer. Put samples on ice. Analyzing samples by flow cytometry using the FL-2 channel. C. Detection by Fluorescence Microscopy: For fluorescence microscopic analysis, resuspend cells in 100 µL Wash buffer. Put one drop of the cell suspension onto a microslide and cover with a coverslip. Observe cells under a fluorescence microscope using rhodamine filter. Caspase-8 positive cells appear to have brighter red signals, whereas caspase-8 negative control cells show much weaker signal. D. Analysis by Fluorescence Plate Reader: For analysis with fluorescence plate reader, resuspend cells in 100 µL Wash Buffer and transfer the cell suspension to each well of the black microtiter plate. Measure the fluorescence intensity at Ex/Em = 540/570 nm (Note: Ex/Em=488/570 nm will also work, although it's not an optimal wavelength). For control, use wells containing unlabeled cells.
Restrictions For Research Use only
Storage -20 °C
Expiry Date 6-12 months
Product cited in: Alajez, Mocanu, Shi, Chia, Breitbach, Hui, Knowles, Bell, Busson, Takada, Lo, OSullivan, Gullane, Liu: "Efficacy of systemically administered mutant vesicular stomatitis virus (VSVDelta51) combined with radiation for nasopharyngeal carcinoma." in: Clinical cancer research : an official journal of the American Association for Cancer Research, Vol. 14, Issue 15, pp. 4891-7, 2008 (PubMed).

Cohen, Raupachova, Wimmer, Deicher, Hörl: "The uraemic retention solute para-hydroxy-hippuric acid attenuates apoptosis of polymorphonuclear leukocytes from healthy subjects but not from haemodialysis patients." in: Nephrology, dialysis, transplantation : official publication of the European Dialysis and Transplant Association - European Renal Association, Vol. 23, Issue 8, pp. 2512-9, 2008 (PubMed).

Moresi, Pristerà, Scicchitano, Molinaro, Teodori, Sassoon, Adamo, Coletti: "Tumor necrosis factor-alpha inhibition of skeletal muscle regeneration is mediated by a caspase-dependent stem cell response." in: Stem cells (Dayton, Ohio), Vol. 26, Issue 4, pp. 997-1008, 2008 (PubMed).

Lin, Lee, Hung, Li, Yang-Yen, Yen: "Survival factor withdrawal-induced apoptosis of TF-1 cells involves a TRB2-Mcl-1 axis-dependent pathway." in: The Journal of biological chemistry, Vol. 282, Issue 30, pp. 21962-72, 2007 (PubMed).

Abdel-Latif, Murray, Renberg, ONeill, Porter, Jensen, Johnson: "Cell death in bovine parvovirus-infected embryonic bovine tracheal cells is mediated by necrosis rather than apoptosis." in: The Journal of general virology, Vol. 87, Issue Pt 9, pp. 2539-48, 2006 (PubMed).

Yip, Ito, Mao, Au, Hedley, Mocanu, Bastianutto, Schimmer, Liu: "Potential use of alexidine dihydrochloride as an apoptosis-promoting anticancer agent." in: Molecular cancer therapeutics, Vol. 5, Issue 9, pp. 2234-40, 2006 (PubMed).