| BML-SE168-5000 | 5000 U | 554.00 USD |  |
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| Alternative Name: | Interleukin 1β converting enzyme, IL-1β converting enzyme, ICE |
| MW: | ~20 + 10kDa |
| Source: | Produced in E. coli. cDNA encodes residues identical to Asn120-His404 (C-terminus) at Genbank Accession No. M87507, except for an Asp381 to Glu change, introduced to stabilize the enzyme against autoproteolysis. |
| UniProt ID: | P29466 |
| Formulation: | Liquid. In 50mM HEPES, pH 7.4, containing 100mM sodium chloride, 0.5% CHAPS, 1mM EDTA, 10% glycerol and 10mM DTT. |
| Purity: | ≥90% (SDS-PAGE) |
| Purity Detail: | Purified by multi-step chromatography. |
| Activity: | 100 U/µl |
| Specific Activity: | One U=1 pmol/min, using Ac-YVAD-pNA (200µM; Prod. No. ALX-260-026) as substrate, at 30°C. |
| Application Notes: | Useful tool to study enzyme regulation and kinetics, cleave target substrates, screen for inhibitors. |
| Shipping: | Dry Ice |
| Long Term Storage: | -80°C |
| Use/Stability: | After initial defrost, aliquot product into individual tubes and refreeze at -80°C. Avoid repeated freeze/defrost cycles.The enzyme is stable on ice for the time typically required to set up an experiment (30-60 min.), but may lose activity with prolonged storage on ice. It is recommended that thawing and dilution of the enzyme be done within as short a time as possible before start of the assay. The remaining, undiluted and unused enzyme should be refrozen quickly by, for example, snap-freezing in a dry ice ethanol bath or liquid nitrogen. The enzyme is stable to at least 4 freeze/thaw cycles. |
| Scientific Background: | First of the caspases described, caspase-1 was subsequently found to be homologous to Ced-3, the C. elegans caspase essential for apoptosis. |
| Protocol: | Caspase-1 (ICE) Assay (Catalog #BML-SE168): Assay buffer: (50mM HEPES, pH 7.4, 100mM NaCl, 0.1% CHAPS, 1mM EDTA, 10% glycerol, 10mM DTT) Caspase-1 (BML-SE168): Dilute to 10 U/μl in assay buffer, just before use. Ac-YVAD-pNA (ALX-260-026) colorimetric substrate: (400 μM stock solution in Assay buffer; Store at -20°C. Warm to assay temperature before use.) To 5 mg net peptide (MW=629) add 159 μl DMSO, to prepare 50 mM stock. Dilute 50mM stock to 400 μM with Assay Buffer. Reaction Conditions: 1) Add 45μl Assay buffer into 1/2 volume microtiter plate. Allow to equilibrate to assay temperature. 2) Add 5μl of caspase-1 (10U/μl) to each appropriate well. Include 2 blanks (5μl assay buffer rather than caspase-1). 3) To start reaction, add 50μl Ac-YVAD-pNA substrate (400μM in assay buffer). Final substrate concentration=200μM. 4) Continuously monitor A405nm. 5) Data analysis: Graph OD vs time and determine slope over the linear portion of the curve. Convert rates in OD/min to substrate/min using an extinction coefficient for pnitroaniline of 10,500M-1 cm-1, and adjusting for pathlength of sample (~0.5cm for 100μl in well of a ½ volume, 96-well plate). |
| Regulatory Status: | RUO - Research Use Only |
Ac-YVAD-pNA |
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| Caspase-1 substrate | ||||||
| 149231-66-3, ≥97% (HPLC) | Print as PDF | ||||||
| ALX-260-026-M001 | 1 mg | 58.00 USD | |
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| ALX-260-026-M005 | 5 mg | 113.00 USD | |
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Ac-YVAD-AMC |
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| Caspase-1 substrate | ||||||
| ≥98% (HPLC) | Print as PDF | ||||||
| ALX-260-024-M001 | 1 mg | 63.00 USD | |
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| ALX-260-024-M005 | 5 mg | 162.00 USD | |
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Caspase-1 assay kit for drug discovery |
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| Colorimetric detection, Fluorescent detection, HTS, Activity assay | Print as PDF | ||||||
| BML-AK701-0001 | 96 wells | 708.00 USD | |
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