[Kamiya Biomedical] Biotin-FA-FMK (Cathepsin B Inhibitor)

[Kamiya Biomedical] Biotin-FA-FMK (Cathepsin B Inhibitor)

 

 

 

Cat. No.: AD-012 (5 mg)

Chemical Name:
Biotin-Phe-Ala-CH2F

Molecular Weight:
478
Description:

Trifluoroacetic acid salt of the fluoromethyl
ketone biotinylated peptide inhibitor of Cathepsin
B (Caspase inhibitor negative control).
The CH2F (fluoromethyl ketone) inhibitor has
several advantages over other types of
derivatives:

• Penetrates cell membranes
• Not toxic to cells
• Irreversible inhibition
• 
  Introduction:
  The ten known caspases are involved in the
  proteolytic cascade that results in apoptosis and
  in the maturation process of IL-1ß necessary for
  inflammation. The use of biotinylated FMK-type
  inhibitors for various caspases requires a
  biotinylated FMK-type inhibitor that does not
  affect caspases as a negative control. The
  Biotin-FA-FMK does not inhibit the activity of
  caspases in intact cells and does not block the
  induction of apoptosis as caspase inhibitors do.
  It can, therefore, be used as a negative control
  for the FMK moiety of the biotinylated caspase
  inhibitor, both in vivo and in vitro.
• 
  Specificity:
  Specifically binds to and inhibits Cathepsin B.
  Does not bind to or inhibit any of the caspases.
  Form:
  Brown solid.
• 
  Applications:
• Irreversible binding to and inhibition of
  Cathepsin B. The second order rate constant (k2)
  of inactivation for human liver Cathepsin B is
  1,220 M-1s
  -1, determined from the observed rate
  of inactivation (k) divided by the inhibitor
  concentration (I): k2 = k/I.
  Serves as a negative control in studies using the
  biotinylated FMK-containing caspase inhibitors.
  Protocol:
  Dissolve Biotin-FA-FMK in high purity (>99.9%)
  DMSO before use.
  For use on intact cells:

1. Prepare desired concentrated stock solution
   in DMSO as follows:
   Dissolve 1 mg Biotin-FA-FMK
   in 68 µL DMSO = 25 mM
   in 85 µL DMSO = 20 mM
   in 169 µL DMSO = 10 mM, etc.
2. Add 2 µL of the stock solution to 1 mL culture
   medium containing cells such that the final
   DMSO concentration is 0.2%. Levels of
   DMSO above this may cause some cellular
   toxicity, thus masking the true effects of the
   inhibitor. The recommended final
   concentration of Biotin-FA-FMK for use as a
   caspase inhibitor negative control is 50 µM (2
   µL of 25 mM stock solution in 1 mL cell
   culture).
3. Detect bound Biotin-FA-FMK by any of
   several avidin/streptavidin methods.
   For extended use in vivo or in vitro:
   For experiments extending 12 to 48 hours, fresh
   inhibitor may have to be added (injected) due to
   inactivation of the inhibitor by endogenous
   cysteine proteases.
   Storage and Stability:
   Store Biotin-FA-FMK in a desiccator at room
   temperature (RT) or 4°C. Biotin-FA-FMK has a
   shelf-life of >2 years if stored desiccated at –
   20°C. For long term, storage at -20°C is
   recommended.
   Limitations:
   For research use only. Not for use in
   diagnostics or in humans.
   Warranty:
   No warranties, expressed or implied, are made
   regarding the use of this product. KAMIYA
   BIOMEDICAL COMPANY is not liable for any
   damage, personal injury