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Recombinant Human HTRA2/Omi Protein, CF

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Summary
Reactivity HuSpecies Glossary
Applications Enzyme Activity
Format
Carrier-Free

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Recombinant Human HTRA2/Omi Protein, CF Summary

Details of Functionality
Measured by its ability to cleave beta -casein. >95% cleavage of beta -casein, as measured under the described conditions.
Source
E. coli-derived human HTRA2/Omi protein
Ala134-Glu458, with a C-terminal 6-His tag
Accession #
N-terminal Sequence
Ala134
Protein/Peptide Type
Recombinant Enzymes
Gene
HTRA2
Purity
>85%, by SDS-PAGE under reducing conditions and visualized by silver stain

Applications/Dilutions

Dilutions
  • Enzyme Activity
Theoretical MW
36 kDa.
Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.
SDS-PAGE
38 kDa, reducing conditions
Publications
Read Publications using
1458-HT in the following applications:

Packaging, Storage & Formulations

Storage
Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 6 months from date of receipt, -20 to -70 °C as supplied.
  • 3 months, -20 to -70 °C under sterile conditions after opening.
Buffer
Supplied as a 0.2 μm filtered solution in HEPES, NaCl, DTT and Glycerol.
Purity
>85%, by SDS-PAGE under reducing conditions and visualized by silver stain
Assay Procedure
  • Assay Buffer: 50 mM Tris, pH 8.0
  • Recombinant Human HTRA2/Omi (rhHTRA2) (Catalog # 1458-HT)
  • Substrate: beta -Casein (Sigma, Catalog # C-6905), 1.0 mg/mL stock in 25 mM Tris, 0.15 M NaCl, pH 7.5
  • SDS-PAGE and silver staining reagents
  1. Dilute Substrate to 0.4 mg/mL in Assay Buffer.
  2. Dilute rhHTRA2 to 0.04 mg/mL in Assay Buffer.
  3. Add 25 μL of substrate and 25 μL of rhHTRA2 to reaction tube. Include a control with 25 μL Substrate and 25 μL Assay Buffer.
  4. Incubate the reaction and control for 1 hour at 45 °C.
  5. Stop the reaction by adding SDS-PAGE sample buffer.
  6. Analyze the cleavage by SDS-PAGE followed by silver staining.
Per Reaction:
  • rhHTRA2: 0.02 mg/mL
  • Substrate: 0.2 mg/mL

Notes

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for Recombinant Human HTRA2/Omi Protein, CF

  • High temperature requirement protein A2
  • HtrA serine peptidase 2
  • HTRA2
  • HtrA-like serine protease
  • Omi
  • OMIOmi stress-regulated endoprotease
  • PARK13EC 3.4.21.108
  • PRSS25
  • PRSS25serine, 25
  • Serine protease 25
  • serine protease HTRA2, mitochondrial
  • Serine proteinase OMI

Background

HtrA2/Omi is the mammalian homologue of bacterial high temperature requirement protein (HtrA). HtrA2/Omi localizes to the mitochondria and is processed to expose an amino-terminal Reaper-like motif similar to SMAC/Diablo. HtrA2/Omi is released from the mitochondria in response to apoptotic insult and can interact with the BIR2 or BIR3 domains of XIAP to relieve caspase-IAP inhibition. This effect can be measured by reversing XIAP-BIR2 (Catalog # 786-XB) inhibition of Caspase-7 (Catalog # 823-C7) cleavage of a fluorogenic peptide (DEVD-AFC, MP Bio, Catalog # AFC-138). IC50 values for this effect are typically between 0.2 and 1.5 μM. HtrA2/Omi is trimeric and functions as a serine protease. The serine protease activity may play a more central role in apoptosis than its IAP antagonizing function. A PDZ domain regulates the serine protease activity by blocking access to the active site. The specificity of the protease is yet to be defined and no endogenous substrates are known to date.

  1. Suzuki, Y. et al. (2001) Mol. Cell. 8:613.
  2. van Loo, G. et al. (2002) Cell Death & Diff. 9:20.
  3. Hedge, R. et al. (2001) J. Biol. Chem. 277:432.
  4. Verhagen, A. et al. (2001) J. Biol. Chem. 277:445.
  5. Martins, L. et al. (2002) J. Biol. Chem. 277:439.
  6. Silke, J., and A. Verhagen (2002) Cell Death & Diff. 9:362.
  7. Savopoulos, J. et al. (2000) Protein Expression & Purification 19:227.

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Recombinant Human
HTRA2/Omi Protein, CF
1458-HT
Recombinant Human HTRA2/Omi Protein, CF
Species: Hu
Applications: Enzyme Activity

Publications for HTRA2/Omi (1458-HT)(5)

We have publications tested in 1 confirmed species: Human.

We have publications tested in 3 applications: Bioassay, Enzyme Assay, Western Blot.


Filter By Application
Bioassay
(1)
Enzyme Assay
(3)
Western Blot
(1)
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Human
(3)
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Showing Publications 1 - 5 of 5.
Publications using 1458-HT Applications Species
G Kaur, D Stallmann, N Schanze, R Laumann, LA Heger, J Steinfurt, P Stachon, K Peter, C Bode, M Moser, I Ahrens, D Duerschmie, M Hortmann Extracellular HtrA2 Induces Apoptosis in Human Umbilical Vein Endothelial Cells Int J Mol Sci, 2019-10-31;20(21):. 2019-10-31 [PMID: 31683713] (Bioassay, Human) Bioassay Human
Singh , Harmeet, Nero , Tracy L, Wang , Yao, Parker , Michael, Nie , Guiying Activity-modulating monoclonal antibodies to the human serine protease HtrA3 provide novel insights into regulating HtrA proteolytic activities. PLoS ONE, 2014-09-23;9(9):e108235. 2014-09-23 [PMID: 25248123] (Western Blot) Western Blot
Chauhan D, Tian Z, Zhou B, Kuhn D, Orlowski R, Raje N, Richardson P, Anderson KC In Vitro and In Vivo Selective Antitumor Activity of a Novel Orally Bioavailable Proteasome Inhibitor MLN9708 against Multiple Myeloma Cells. Clin. Cancer Res., 2011-06-30;17(16):5311-21. 2011-06-30 [PMID: 21724551] (Enzyme Assay) Enzyme Assay
Johnson F, Kaplitt MG Novel mitochondrial substrates of omi indicate a new regulatory role in neurodegenerative disorders. PLoS ONE, 2009-09-18;4(9):e7100. 2009-09-18 [PMID: 19763263] (Enzyme Assay, Human) Enzyme Assay Human
Huttunen HJ, Guenette SY, Peach C, Greco C, Xia W, Kim DY, Barren C, Tanzi RE, Kovacs DM HtrA2 regulates beta-amyloid precursor protein (APP) metabolism through endoplasmic reticulum-associated degradation. J. Biol. Chem., 2007-08-06;282(38):28285-95. 2007-08-06 [PMID: 17684015] (Enzyme Assay, Human) Enzyme Assay Human

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Bioinformatics

Gene Symbol HTRA2
Uniprot