CaspGLOWTM Fluorescein Active Caspase-12 Staining Kit

Details for Product No. ABIN411813, Supplier: Log in to see
Antigen
  • Caspase-12
  • caspase 12
  • Casp12
  • CASP12
Alternatives
Human Caspase 12 ELISA Kit
Reactivity
Mammalian
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Application
Detection (D), Fluorescence Microscopy (FM), Flow Cytometry (FACS)
Options
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Purpose The CaspGLOW™ Fluorescein Caspase-12 Staining Kit provides a convenient means for sensitive detection of activated caspase-12 in living cells. The assay utilizes the caspase-12 inhibitor, ATAD-FMK, conjugated to FITC (FITC-ATAD-FMK) as a marker. FITC-ATAD-FMK is cell permeable, nontoxic, and irreversibly binds to activated caspase-12 in apoptotic cells.
Brand CaspGLOW™
Sample Type Cell Samples
Detection Method Fluorometric
Specificity The CaspGLOW™ Fluorescein Caspase-12 Staining Kit provides a convenient means for sensitive detection of activated caspase-12 in living cells. The assay utilizes the caspase-12 inhibitor, ATAD-FMK, conjugated to FITC (FITC-ATAD-FMK) as a marker. FITC-ATAD-FMK is cell permeable, nontoxic, and irreversibly binds to activated caspase-12 in apoptotic cells. The FITC label allows for direct detection of activated caspases in apoptotic cells by fluorescence microscopy, flow cytometry, or fluorescence plate reader.
Characteristics CaspGLOWTM Fluorescein Active Caspase-12 Staining Kit: Convenient & Sensitive Kit to Detect Activated Caspase-12 in Living Cells. Detection Method: Fluorescence Microscopy, Flow Cytometry or Fluorescence Plate Reader.
Components FITC-ATAD-FMK
Wash Buffer
Z-VAD-FMK
Alternative Name Caspase-12 (CASP12 ELISA Kit Abstract)
Background Activation of caspases plays a central role in apoptosis. The CaspGLOW™ Fluorescein Caspase-12 Staining Kit provides a convenient means for sensitive detection of activated caspase-12 in living cells. The assay utilizes the caspase-12 inhibitor, ATAD-FMK, conjugated to FITC (FITC-ATAD-FMK) as a marker. FITC-ATAD-FMK is cell permeable, nontoxic, and irreversibly binds to activated caspase-12 in apoptotic cells.
Research Area Apoptosis/Necrosis
Application Notes Sensitive detection of activated caspase-12 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-12 activation.
2. Aliquot 300 µL each of the induced and control cultures into eppendorf tubes.
3. Add 1 µL of FITC-ATAD-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. Keep samples on ice. Analyzing samples by flow cytometry using the FL-1 channel. C. Detection by Fluorescence Microscopy: For fluorescence microscopic analysis, resuspend cells in 100 µL Wash Buffer. Transfer one drop of the cell suspension onto a microslide and cover with a coverslip. Observe cells under a fluorescence microscope using FITC filter. Caspase positive cells appear to have brighter green signals, whereas caspase 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 then transfer the cell suspension into each well in the black microtiter plate. Measure the fluorescence intensity at Ex. = 485 nm and Em. = 535 nm. For control, use wells containing unlabeled cells.
Restrictions For Research Use only
Storage -20 °C
Expiry Date 6-12 months
Product cited in: Su, Wu, Hong: "Betanodavirus up-regulates chaperone GRP78 via ER stress: roles of GRP78 in viral replication and host mitochondria-mediated cell death." in: Apoptosis : an international journal on programmed cell death, Vol. 16, Issue 3, pp. 272-87, 2011 (PubMed).

Dolai, Pal, Yadav, Adak: "Endoplasmic reticulum stress-induced apoptosis in Leishmania through Ca2+-dependent and caspase-independent mechanism." in: The Journal of biological chemistry, Vol. 286, Issue 15, pp. 13638-46, 2011 (PubMed).

Auner, Beham-Schmid, Dillon, Sabbattini: "The life span of short-lived plasma cells is partly determined by a block on activation of apoptotic caspases acting in combination with endoplasmic reticulum stress." in: Blood, Vol. 116, Issue 18, pp. 3445-55, 2010 (PubMed).

Cunha, Ladrière, Ortis, Igoillo-Esteve, Gurzov, Lupi, Marchetti, Eizirik, Cnop: "Glucagon-like peptide-1 agonists protect pancreatic beta-cells from lipotoxic endoplasmic reticulum stress through upregulation of BiP and JunB." in: Diabetes, Vol. 58, Issue 12, pp. 2851-62, 2009 (PubMed).

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).

Chwa, Atilano, Hertzog, Zheng, Langberg, Kim, Kenney: "Hypersensitive response to oxidative stress in keratoconus corneal fibroblasts." in: Investigative ophthalmology & visual science, Vol. 49, Issue 10, pp. 4361-9, 2008 (PubMed).

Bian, Elner, Elner: "Regulated expression of caspase-12 gene in human retinal pigment epithelial cells suggests its immunomodulating role." in: Investigative ophthalmology & visual science, Vol. 49, Issue 12, pp. 5593-601, 2008 (PubMed).

Trisciuoglio, Uranchimeg, Cardellina, Meragelman, Matsunaga, Fusetani, Del Bufalo, Shoemaker, Melillo: "Induction of apoptosis in human cancer cells by candidaspongiolide, a novel sponge polyketide." in: Journal of the National Cancer Institute, Vol. 100, Issue 17, pp. 1233-46, 2008 (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).

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