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Recombinant Human HTRA2/Omi Protein, CF

Catalog #: 1458-HT
6 Citations Datasheet / COA / SDS
Catalog # Availability Size / Price Qty
1458-HT-100
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Citations (6)
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Summary Product Datasheets Carrier Free Data Images Reconstitution Calculator Background Related Research Areas

Recombinant Human HTRA2/Omi Protein, CF Summary

Product Specifications

Purity
>85%, by SDS-PAGE under reducing conditions and visualized by silver stain
Activity
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 #
O43464
N-terminal Sequence
Analysis
Ala134
Predicted Molecular Mass
36 kDa
SDS-PAGE
38 kDa, reducing conditions

Product Datasheets

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1458-HT

Product Datasheet
COA
SDS

Carrier Free

What does CF mean?

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.

What formulation is right for me?

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.
  • 6 months from date of receipt, -20 to -70 °C as supplied.
  • 3 months, -20 to -70 °C under sterile conditions after opening.

Assay Procedure

Materials
  • 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
Reconstitution Calculator

Reconstitution Calculator

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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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.

References
  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.
Long Name
High Temperature Requirement Protein-2
Entrez Gene IDs
27429 (Human); 64704 (Mouse)
Alternate Names
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

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.

6 Citations: Showing 1 - 6
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  1. 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)
  2. 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
  3. 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
  4. 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
  5. 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
  6. 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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