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96-Well Plate
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48-602MAG
Buffer Detection Kit for Magnetic Beads
1 Kit
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CC120
Sigma-AldrichCaspase 9, recombinant human active
Caspase 9, recombinant human active: SDB (Sicherheitsdatenblätter), Analysenzertifikate und Qualitätszertifikate, Dossiers, Broschüren und andere verfügbare Dokumente.
Caspase-9 is a member of the interleukin-1beta converting enzyme (ICE) family of cysteine proteases. Similar to other caspases, caspase-9 also exists in cells as an inactive proenzyme. During the initiation of apoptosis the proenzyme is processed at aspartate residues to form active caspase-9. As one of the initiator caspases, active caspase-9 functions to trigger the activation of downstream effector caspases and further lead to disassembly of cell structures.
The partially purified active recombinant human caspase-9 with full-length HIS-6 tag was expressed in E. coli. The expressed caspase-9 spontaneously undergoes autoprocessing to yield the subunits characteristic of the native enzyme. In combination with caspase-9 activity assays, the active recombinant caspase-9 is useful in biological screening of caspase inhibitors. The recombinant enzyme can also be used as a positive control in caspase-9 assays. Caspase 9 is known to have strong reactivity with other sequence peptides, especially DEVD-pNA.
Alternate Names
APAF-3
ICE-LAP6
Mch-6
References
Product Information
Presentation
Lyophilized. Reconstitute with 25 μl of PBS + 15% glycerol.
Enzyme is prepared as an activity standard only, any other application not tested. Caspase-9 is subject to auto-activation thus active caspase subunits and partially active intermediates will be seen. Bands of approximately 45kDa (pro), 35kDa (partially active) and 18kDa + 12kDa (large and small subunits) may be seen by western blot depending upon the antibody used. The small subunit is extremely difficult to detect in most westerns.
Activation: incubate 1-5 μL of stock in PBS pH 7.5 (100mM HEPES can be used as well) plus 10mM ATP and 5mM DTT (added just before use) for 30 minutes at 37°C.
Biological Information
Purity
10-20% caspase-9 protein by gel analysis. Also contains E. coli proteins. Approximately, 0.8-2 μg total protein / unit of activity.
Source
Human
Specific Activity
One unit of the recombinant caspase-9 is the enzyme activity that cleaves 1 nmol of the caspase substrate LEHD-pNA (pNA: pnitroanaline) per hour at 37°C at saturated substrate concentrations.
This gene encodes a member of the cysteine-aspartic acid protease (caspase) family. Sequential activation of caspases plays a central role in the execution-phase of cell apoptosis. Caspases exist as inactive proenzymes which undergo proteolytic processing at conserved aspartic residues to produce two subunits, large and small, that dimerize to form the active enzyme. This protein is processed by caspase APAF1; this step is thought to be one of the earliest in the caspase activation cascade. Alternative splicing results in two transcript variants which encode different isoforms.
FUNCTION: SwissProt: P55211 # Isoform 2 lacks activity is an dominant-negative inhibitor of caspase-9. SIZE: 416 amino acids; 46281 Da SUBUNIT: Heterotetramer that consists of two anti-parallel arranged heterodimers, each one formed by a 35 kDa (p35) and a 10 kDa (p10) subunit. Caspase-9 and APAF1 bind to each other via their respective NH2-terminal CED-3 homologous domains in the presence of cytochrome C and ATP. Interacts with the inhibitors BIRC2, BIRC4, BIRC5 and BIRC7. TISSUE SPECIFICITY: Ubiquitous, with highest expression in the heart, moderate expression in liver, skeletal muscle, and pancreas. Low levels in all other tissues. PTM: Cleavages at Asp-315 by granzyme B and at Asp-330 by caspase- 3 generate the two active subunits. Caspase-8 and -10 can also be involved in these processing events. SIMILARITY: SwissProt: P55211 ## Belongs to the peptidase C14 family. & Contains 1 CARD domain.
Physicochemical Information
Dimensions
Materials Information
Toxicological Information
Safety Information according to GHS
Safety Information
Product Usage Statements
Usage Statement
Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.
Storage and Shipping Information
Storage Conditions
Maintain at -70°C in undiluted aliquots for 3-6 months from date of receipt. Avoid repeated freeze/thaw cycles.
Neuroprotective effect of hyperbaric oxygen therapy in brain injury is mediated by preservation of mitochondrial membrane properties. Eilam Palzur,Menashe Zaaroor,Eugene Vlodavsky,Felix Milman,Jean F Soustiel Brain research
1221
2008
Recent experimental data have shown that hyperbaric oxygen therapy (HBOT) was associated increased Bcl-2 expression at the injury site that correlated with reduced apoptosis. We hypothesized that HBOT mediated enhancement of Bcl-2 expression and increased intracellular oxygen bio-availability may both contribute to preserve mitochondrial integrity and reduce the activation of the mitochondrial pathway of apoptosis. For this purpose, a cortical lesion was created in the parietal cortex of Sprague-Dawley rats by dynamic cortical deformation (DCD) and outcome measures in non-treated animals were compared with that of HBOT treated rats. Morphological analysis showed a profound reduction in neuronal counts in the perilesional area and a marked rarefaction of the density of the axonal-dendritic network. In treated animals, however, there was a significant attenuation of the impact of DCD over perilesional neurons, characterized by significantly higher cell counts and denser axonal network. In mitochondria isolated from injured brain tissue, there was a profound loss of mitochondrial transmembrane potential (Deltapsi(M)) that proved to be substantially reversed by HBOT. This finding correlated with a significant reduction of caspases 3 and 9 activation in HBOT treated animals but not of caspase 8, indicating a selective effect over the intrinsic pathway of apoptosis. All together, our results indicate that the neuroprotective effect of HBOT may represent the consequence of preserved mitochondrial integrity and subsequent inhibition of the mPTP and reduction of the mitochondrial pathway of apoptosis.
A conserved coronavirus epitope, critical in virus neutralization, mimicked by internal-image monoclonal anti-idiotypic antibodies. C Suñé,C Smerdou,I M Antón,P Abril,J Plana,L Enjuanes Journal of virology
65
1991
Monoclonal antibody (MAb) 6A.C3 neutralizes transmissible gastroenteritis coronavirus (TGEV) and is specific for a conserved epitope within subsite Ac of the spike (S) glycoprotein of TGEV. Six hybridomas secreting anti-idiotypic (Ab2) MAbs specific for MAb 6A.C3 (Ab1) have been selected. All six MAbs inhibited the binding of Ab1 to TGEV and specifically cross-linked MAb1-6A.C3. Four of these hybridomas secreted gamma-type anti-idiotypic MAbs. The other two Ab2s (MAbs 9A.G3 and 9C.E11) were recognized by TGEV-specific antiserum induced in two species. This binding was inhibited by viruses of the TGEV group but not by serologically unrelated coronaviruses. These results indicate that MAb2-9A.G3 and MAb2-9C.E11 mimic an antigenic determinant present on the TGEV surface, and they were classified as beta-type (internal-image) MAbs. TGEV-binding Ab3 antiserum was induced in 100% of mice immunized with the two beta-type MAb2s and in 25 to 50% of mice immunized with gamma-type MAb2. Both beta- and gamma-type Ab2s induced neutralizing Ab3 antibodies in mice that were mainly directed to antigenic subsite Ac of the S protein.
Millipore’s Caspase Antibodies, proteins, and assays have been well validated and published. See below for a comprehensive list of our Caspase products, based on the expertise of Upstate & Chemicon. Weitere Informationen >>
