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Caspase-1 (human), (recombinant)

BML-SE168

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Highly active caspase essential for apoptosis.

First of the caspases described, caspase-1 was subsequently found to be homologous to Ced-3, the C. elegans caspase essential for apoptosis.

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Caspase-1 (human), (recombinant) SDS-PAGE

Product Details

Activity	100 U/µl
Alternative Name	Interleukin 1β converting enzyme, IL-1β converting enzyme, ICE
Application Notes	Useful tool to study enzyme regulation and kinetics, cleave target substrates, screen for inhibitors.
Formulation	Liquid. In 50mM HEPES, pH 7.4, containing 100mM sodium chloride, 0.5% CHAPS, 1mM EDTA, 10% glycerol and 10mM DTT.
MW	~20 + 10kDa
Purity	≥90% (SDS-PAGE)
Purity Detail	Purified by multi-step chromatography.
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.
Specific Activity	One U=1 pmol/min, using Ac-YVAD-pNA (200µM; Prod. No. ALX-260-026) as substrate, at 30°C.
UniProt ID	P29466

Handling & Storage

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.
Long Term Storage	-80°C
Shipping	Dry Ice

Regulatory Status	RUO – Research Use Only

A Potent Inhibitor of Caspase‑8 Based on the IL-18 Tetrapeptide Sequence Reveals Shared Specificities between Inflammatory and Apoptotic Initiator Caspases: C.M. Bourne, et al.; bioRxiv , 02.23.639785 (2025), Abstract — Full Text
The tetrapeptide sequence of IL-18 and IL-1β regulates their recruitment and activation by inflammatory caspases: Exconde, P. M., Hernandez-Chavez, C., et al.; Cell Rep. 42, 113581 (2023), Abstract
A small molecule inhibitor of caspase-1 inhibits NLRP3 inflammasome activation and pyroptosis to alleviate gouty inflammation: D.Y. Cao, et al.; Immunol. Lett. 244, 28 (2022), Abstract
The colonic pathogen Entamoeba histolytica activates caspase-4/1 that cleaves the pore-forming protein gasdermin D to regulate IL-1β secretion: S. Wang, et al.; PLoS Pathog. 18, e1010415 (2022), Abstract
An Essential Role for SHARPIN in the Regulation of Caspase 1 Activity in Sepsis: M.V. Nastase, et al.; Am. J. Pathol. 186, 1206 (2016), Abstract
iGLuc: a luciferase-based inflammasome and protease activity reporter: E. Bartok, et al.; Nat. Methods 10, 147 (2013), Application(s): Recombinant Proteolysis, Abstract — Full Text
The Yersinia virulence effector YopM binds caspase-1 to arrest inflammasome assembly and processing: C.N. LaRock, et al.; Cell Host Microbe 12, 799 (2012), Application(s): Incubation, Abstract — Full Text
Antiapoptotic mechanism of HIV protease inhibitors: preventing mitochondrial transmembrane potential loss: N. Barbara, et al.; Blood 98, 1078 (2001), Abstract — Full Text
Preparation of an autolysis-resistant interleukin-1 beta converting enzyme mutant: L.C. Dang, et al.; Biochemistry 35, 14910 (1996), Abstract
Interleukin-1 beta converting enzyme: N.A. Thornberry; Methods Enzymol. 244, 615 (1994), Abstract

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Datasheet, Manuals, SDS & CofA

Manuals And Inserts

Datasheet

Specific 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).

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