Recombinant Human HTRA2/Omi Protein, CF Summary
Product Specifications
Ala134-Glu458, with a C-terminal 6-His tag
Analysis
Product Datasheets
1458-HT
Carrier Free
CF stands for Carrier Free (CF). We typically add Bovine Serum Albumin (BSA) as a carrier protein to our recombinant proteins. Adding a carrier protein enhances protein stability, increases shelf-life, and allows the recombinant protein to be stored at a more dilute concentration. The carrier free version does not contain BSA.
In general, we advise purchasing the recombinant protein with BSA for use in cell or tissue culture, or as an ELISA standard. In contrast, the carrier free protein is recommended for applications, in which the presence of BSA could interfere.
1458-HT
| Formulation | Supplied as a 0.2 μm filtered solution in HEPES, NaCl, DTT and Glycerol. |
| Shipping | The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below. |
| Stability & Storage: | Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
|
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
- Dilute Substrate to 0.4 mg/mL in Assay Buffer.
- Dilute rhHTRA2 to 0.04 mg/mL in Assay Buffer.
- 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.
- Incubate the reaction and control for 1 hour at 45 °C.
- Stop the reaction by adding SDS-PAGE sample buffer.
- Analyze the cleavage by SDS-PAGE followed by silver staining.
- rhHTRA2: 0.02 mg/mL
- Substrate: 0.2 mg/mL
Background: HTRA2/Omi
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.
- Suzuki, Y. et al. (2001) Mol. Cell. 8:613.
- van Loo, G. et al. (2002) Cell Death & Diff. 9:20.
- Hedge, R. et al. (2001) J. Biol. Chem. 277:432.
- Verhagen, A. et al. (2001) J. Biol. Chem. 277:445.
- Martins, L. et al. (2002) J. Biol. Chem. 277:439.
- Silke, J., and A. Verhagen (2002) Cell Death & Diff. 9:362.
- Savopoulos, J. et al. (2000) Protein Expression & Purification 19:227.
Citations for Recombinant Human HTRA2/Omi Protein, CF
R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.
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Citations: Showing 1 - 6
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Cystine-knot peptide inhibitors of HTRA1 bind to a cryptic pocket within the active site region
Authors: Li, Y;Wei, Y;Ultsch, M;Li, W;Tang, W;Tombling, B;Gao, X;Dimitrova, Y;Gampe, C;Fuhrmann, J;Zhang, Y;Hannoush, RN;Kirchhofer, D;
Nature communications
Species: N/A
Sample Types: Fluorogenic Peptide Substrate, Peptide, Recombinant Protein
Applications: Bioassay, Surface Plasmon Resonance (SPR) -
Extracellular HtrA2 Induces Apoptosis in Human Umbilical Vein Endothelial Cells
Authors: 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
Int J Mol Sci, 2019-10-31;20(21):.
Species: Human
Sample Types: Whole Cells
Applications: Bioassay -
Activity-modulating monoclonal antibodies to the human serine protease HtrA3 provide novel insights into regulating HtrA proteolytic activities.
Authors: Singh, Harmeet, Nero, Tracy L, Wang, Yao, Parker, Michael, Nie, Guiying
PLoS ONE, 2014-09-23;9(9):e108235.
Applications: Western Blot -
In Vitro and In Vivo Selective Antitumor Activity of a Novel Orally Bioavailable Proteasome Inhibitor MLN9708 against Multiple Myeloma Cells.
Authors: Chauhan D, Tian Z, Zhou B, Kuhn D, Orlowski R, Raje N, Richardson P, Anderson KC
Clin. Cancer Res., 2011-06-30;17(16):5311-21.
Applications: Enzyme Assay -
Novel mitochondrial substrates of omi indicate a new regulatory role in neurodegenerative disorders.
Authors: Johnson F, Kaplitt MG
PLoS ONE, 2009-09-18;4(9):e7100.
Species: Human
Sample Types: Cell Lysates
Applications: Enzyme Assay -
HtrA2 regulates beta-amyloid precursor protein (APP) metabolism through endoplasmic reticulum-associated degradation.
Authors: Huttunen HJ, Guenette SY, Peach C, Greco C, Xia W, Kim DY, Barren C, Tanzi RE, Kovacs DM
J. Biol. Chem., 2007-08-06;282(38):28285-95.
Species: Human
Sample Types: Buffer
Applications: Enzyme Assay
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