產(chǎn)品詳情

  • 產(chǎn)品名稱：Biotin-VAD-FMK(Pan-CaspaseInhibitor)

  • 產(chǎn)品型號(hào)：
  • 產(chǎn)品廠商：KamiyaBiomedical
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Biotin-VAD-FMK(Pan-CaspaseInhibitor)
詳情介紹：
Purpose Irreversible pan-caspase inhibitor?
Sequence Biotin-Val-Ala-Asp(OMe)-CH2F
Specificity Broad-spectrum caspase inhibitor.
Characteristics Dissolve the caspase inhibitor in high purity DMSO (>99.9%) before use to make a stock solution of 20 mM.

Method for assay of Biotin-VAD-FMK Inhibitor:
1. Grow and treat 1 x 10^6 cells at the appropriate dose and time to obtain 50 - 60% apoptosis.
2. Collect cells by centrifugation, remove supernatant and re-suspend cell pellet in an initial volume of 1/1,000 th of the original media volume (10 μL for 10 mL media).
3. Add 20 μL of 2X Biotin-VAD-FMK in MGD buffer. (Final Biotin-VAD-FMK concentration = 10 μM.)
4. Freeze/thaw 3X to lyse cells.
5. Incubate at 37°C for 15 minutes. Spin two minutes at 14K to remove cell debris.
6. Transfer supernatant to new tube, add 13 μL of 4X SDS-PAGE buffer and run all on 10% SDS-PAGE gel and transfer to PVDF membrane.
7. Block one hour and incubate with streptavidin-HRP (1/1,000) for four hours. Develop by ECL.
MGD Buffer:
50 mM NaCl
2 mM MgCl2
5 mM EDTA
10 mM HEPES pH 7.0
1 mM DTT
Protease Inhibitor

IMPORTANT NOTE for in vitro use: Our peptide inhibitors are synthesized as methyl esters to enhance cell permeability. In intact cells, the methyl groups are removed by endogenous enzymes. For in vitro experiments with purified enzymes, however, the methyl groups must first be removed by treating the inhibitor with esterase. A procedure is available upon request.
Permeability Cell-permeable
Solubility DMSO
Molecular Weight 672 Da
Comment

Biotinylated peptide-fluoromethyl ketone inhibitor of caspases. The FMK (fluoromethyl ketone, CH2F) inhibitor has several advantages over other types of derivatives: Penetrates cell membranes, Not toxic to cells, Irreversible inhibition.
Restrictions For Research Use only
Format Solid
Storage 4 °C