Replaces Prod. #: BML-P443
Chromogenic substrate for caspase-9 and cysteine proteases. Similar to Ac-LEHD-AMC (Prod. No. ALX-260-080) and Ac-LEHD-AFC (Prod. No. ALX-260-116) but cleavage is monitored colorimetrically by absorbance at 405nm.
Product Details
Alternative Name: | Caspase-9 substrate (chromogenic) |
|
Sequence: | Ac-Leu-Glu-His-Asp-pNA (pNA = p-Nitroaniline) |
|
Formula: | C29H38N8O11 |
|
MW: | 674.7 |
|
Formulation: | Lyophilized. |
|
Peptide Content: | 75-95% |
|
Purity: | ≥99% (HPLC) |
|
Solubility: | Soluble in water or DMSO. Dilute with buffer, pH 7.5. |
|
Shipping: | Ambient |
|
Long Term Storage: | -20°C |
|
Technical Info/Product Notes: | λmax of pNA is 400nm. |
|
Protocol: | HEPES-Buffer: 100 mM HEPES, pH 7.5; 20% (v/v)
glycerol; 5 mM DTT, 0.5 mM EDTA.
Substrate: Prepare 1 mM stock solution in DMSO.
- Induce apoptosis and prepare cell lysate or use recombinant caspase.
- Prepare reaction solution:
30 µl of substrate stock solution + 240 µl HEPES-Buffer + 30 µl of cell lysate.
- Incubate for 1 hour at 37 °C.
- Measure with microplate reader at 400nm.
- Suggested controls: Reaction mixture…
…without substrate.
…with non-apoptotic cell lysate.
…with apoptotic cell lysate and caspase inhibitor. |
|
Regulatory Status: | RUO - Research Use Only |
|
Product Literature References
Self-assembly of nanomicelles with rationally designed multifunctional building blocks for synergistic chemo-photodynamic therapy: G. Gong, et al.; Theranostics
12, 2028 (2022),
Abstract;
Cardioprotective effects of Prolame and SNAP are related with nitric oxide production and with diminution of caspases and calpain-1 activities in reperfused rat hearts: N.G. Roman-Anguiano, et al.; PeerJ
7, e7348 (2019),
Abstract;
Full Text
The preclinical analysis of TW-37 as a potential anti-colorectal cancer cell agent: S. Lei, et al.; PLoS One
12, e0184501 (2017),
Abstract;
Full Text
Early effects of Aβ1-42 oligomers injection in mice: Involvement of PI3K/Akt/GSK3 and MAPK/ERK1/2 pathways: F. Morroni, et al.; Behav. Brain Res.
314, 106 (2016),
Application(s): Determination of caspase-9 activation in tissue lysates,
Abstract;
A combinatorial approach defines specificities of members of the caspase family and granzyme B. Functional relationships established for key mediators of apoptosis: N.A. Thornberry et al.; J. Biol. Chem.
272, 17907 (1997),
Abstract;
Related Products