Recombinant Human HTRA1 Protein, CF Summary
Product Specifications
Gly156-Pro480, with an N-terminal Met and C-terminal 6-His tag
Analysis
Product Datasheets
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.
2916-SE
| Formulation | Supplied as a 0.2 μm filtered solution in MES and NaCl. |
| 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 HTRA1/PRSS11 (rhHTRA1) (Catalog # 2916-SE)
- beta -Casein (Sigma, Catalog # C6905), 1 mg/mL in 25 mM Tris, 150 mM NaCl, pH 7.5
- Reducing SDS-PAGE Sample Buffer
- SDS-PAGE or Western Blot
- Dilute beta -Casein to 100 µg/mL in Assay Buffer.
- Dilute rhHTRA1 to 10 µg/mL in Assay Buffer.
- In a vial, combine equal volumes of diluted beta -Casein and diluted rhHTRA1. Also prepare two beta -Casein controls containing Assay Buffer in place of rhHTRA1.
- Incubate reaction and one of the controls at 37 °C for 30 minutes. Keep the other control frozen at -20 °C.
- After incubation, combine 25 µL of the reaction and control with 25 µL of reducing SDS-PAGE gel buffer. Mix and heat samples at 95‑100 °C for 3-5 minutes.
- Load 40 µL per lane and analyze the cleavage by SDS-PAGE followed by protein staining and/or Western blot.
- rhHTRA1: 0.1 µg
- beta -Casein: 1 µg
Reconstitution Calculator
Background: HTRA1/PRSS11
HTRA1 is a member of the mammalian HTRA (high temperature requirement A) serine protease family. The E. coli HTRA homolog functions as a chaperone-protease and is essential for bacterial survival at temperatures above 42 °C (1). Among the four mammalian HTRA proteins, HTRA1, -3 and -4 are secreted while HTRA2 is localized in mitochondria. HTRA1 contains an N-terminal insulin-like growth factor binding protein (IGFBP) domain, a Kazal-type trypsin inhibitor motif, and C-terminal trypsin-like protease and PDZ domains (2). It is involved in several pathologies including Alzheimer’s disease (3), rheumatoid arthritis (4), osteoarthritis (5) and age‑related macular degeneration (6), although the mechanisms by which HTRA1 exerts its effects are not clear. HTRA1 also has properties of a tumor suppressor protein, which makes it a target for cancer therapy (7). In addition HTRA1 is known to regulate the TGF-beta signaling pathway and bone mineralization (8, 9). Recombinant human HTRA1 lacks the N-terminal IGFBP and Kazal-like domains, but retains the serine protease and PDZ domains.
- Jiang, J. et al. (2008) Proc. Natl. Acad. Sci. U.S.A. 105:11939.
- Murwantoko et al. (2004) Biochem. J. 381:895.
- Grau, S. et al. (2005) Proc. Natl. Acad. Sci. U.S.A. 102:6021.
- Grau, S. et al. (2006) J. Biol. Chem. 281:6124.
- Hu, S. et al. (1998) J. Biol. Chem. 273:34406.
- Dewan, A. et al. (2006) Science 314:989.
- Chien, J. et al. (2004) Oncogene 23:1636.
- Oka, C. et al. (2004) Development 131:1041.
- Hadfield, K.D. et al. (2008) J. Biol. Chem. 283:5928.
Citations for Recombinant Human HTRA1 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 - 5
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Exploring the Therapeutic Potential of Elastase Inhibition in Age-Related Macular Degeneration in Mouse and Human
Authors: Navneet, S;Brandon, C;Simpson, K;Rohrer, B;
Cells
Species: Human
Sample Types: Whole Cells
Applications: Bioassay -
Cell density-dependent proteolysis by HtrA1 induces translocation of zyxin to the nucleus and increased cell survival
Authors: F Sabino, E Madzharova, U Auf dem Ke
Cell Death Dis, 2020-08-21;11(8):674.
Species: Human
Sample Types: Protein
Applications: Bioassay -
HtrA1 Is Specifically Up-Regulated in Active Keloid Lesions and Stimulates Keloid Development
Authors: S Yamawaki, M Naitoh, H Kubota, R Aya, Y Katayama, T Ishiko, T Tamura, K Yoshikawa, T Enoshiri, M Ikeda, S Suzuki
Int J Mol Sci, 2018-04-24;19(5):.
Species: Human
Sample Types: Whole Cells
Applications: Bioassay -
BMP-responsive Protease HtrA1 is Differentially Expressed in Astrocytes and Regulates Astrocytic Development and Injury Response
Authors: J Chen, S Van Gulden, TL McGuire, AC Fleming, C Oka, JA Kessler, CY Peng
J. Neurosci., 2018-02-26;0(0):.
Species: Mouse
Sample Types: Whole Cells
Applications: Bioassay -
The inhibitory effect of IFN-gamma on protease HTRA1 expression in rheumatoid arthritis.
Authors: Hou Y, Lin H, Zhu L, Liu Z, Hu F, Shi J, Yang T, Shi X, Guo H, Tan X, Zhang L, Wang Q, Li Z, Zhao Y
J Immunol, 2014-06-06;193(1):130-8.
Applications: ELISA (Standard)
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