Recombinant Human HTRA1 Protein, CF

• Reactivity: Hu
• Applications: Enzyme Activity
• Format: Carrier-Free

Recombinant Human HTRA1 Protein, CF Summary

• Details of Functionality: Measured by its ability to cleave beta -casein. >95% of beta -Casein is cleaved by Recombinant Human HTRA1/PRSS11, as measured under the described conditions.
• Source: E. coli-derived human HTRA1/PRSS11 protein
  Gly156-Pro480, with an N-terminal Met and C-terminal 6-His tag
• Accession #: Q92743
• N-terminal Sequence: Met
• Protein/Peptide Type: Recombinant Enzymes
• Gene: HTRA1
• Purity: >90%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining
• Endotoxin Note: <1.0 EU per 1 μg of the protein by the LAL method.

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: 37-40 kDa, reducing conditions

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 MES and NaCl.
• Purity: >90%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining
• 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
  1. Dilute beta -Casein to 100 µg/mL in Assay Buffer.
  2. Dilute rhHTRA1 to 10 µg/mL in Assay Buffer.
  3. 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.
  4. Incubate reaction and one of the controls at 37 °C for 30 minutes. Keep the other control frozen at -20 °C.
  5. 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.
  6. Load 40 µL per lane and analyze the cleavage by SDS-PAGE followed by protein staining and/or Western blot.
  Per Lane:
  • rhHTRA1: 0.1 µg
  • beta -Casein: 1 µg

Notes

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

Alternate Names for Recombinant Human HTRA1 Protein, CF

• ARMD7
• EC 3.4.21
• EC 3.4.21.108
• High-temperature requirement A serine peptidase 1
• HtrA serine peptidase 1
• HTRA1
• IGFBP5-protease
• L56
• ORF480
• PRSS11
• Serine protease 11
• serine, 11 (IGF binding)

Background

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.

1. Jiang, J. et al. (2008) Proc. Natl. Acad. Sci. U.S.A. 105:11939.
2. Murwantoko et al. (2004) Biochem. J. 381:895.
3. Grau, S. et al. (2005) Proc. Natl. Acad. Sci. U.S.A. 102:6021.
4. Grau, S. et al. (2006) J. Biol. Chem. 281:6124.
5. Hu, S. et al. (1998) J. Biol. Chem. 273:34406.
6. Dewan, A. et al. (2006) Science 314:989.
7. Chien, J. et al. (2004) Oncogene 23:1636.
8. Oka, C. et al. (2004) Development 131:1041.
9. Hadfield, K.D. et al. (2008) J. Biol. Chem. 283:5928.

Publications for HTRA1/PRSS11 (2916-SE)(6)

Publications using 2916-SE

• Aapjeet, F;Tang, T;Zhang, Y;Gopalan, A;Kallappagoudar, S;Pan, J;Ma, F;Shah, SS;Rivera, S;Liu, AP;Juan, V;Liu, R; Fragmentation of recombinant human interleukin-12 by matriptase in CHO cell culture Journal of biotechnology 2025-04-18 [PMID: 40254034] (Bioassay, N/A)
• Navneet, S;Brandon, C;Simpson, K;Rohrer, B; Exploring the Therapeutic Potential of Elastase Inhibition in Age-Related Macular Degeneration in Mouse and Human Cells 2023-05-03 [PMID: 37174708] (Bioassay, Human)
• F Sabino, E Madzharova, U Auf dem Ke Cell density-dependent proteolysis by HtrA1 induces translocation of zyxin to the nucleus and increased cell survival Cell Death Dis, 2020-08-21;11(8):674. 2020-08-21 [PMID: 32826880] (Bioassay, Human)
• S Yamawaki, M Naitoh, H Kubota, R Aya, Y Katayama, T Ishiko, T Tamura, K Yoshikawa, T Enoshiri, M Ikeda, S Suzuki HtrA1 Is Specifically Up-Regulated in Active Keloid Lesions and Stimulates Keloid Development Int J Mol Sci, 2018-04-24;19(5):. 2018-04-24 [PMID: 29695130] (Bioassay, Human)
• J Chen, S Van Gulden, TL McGuire, AC Fleming, C Oka, JA Kessler, CY Peng BMP-responsive Protease HtrA1 is Differentially Expressed in Astrocytes and Regulates Astrocytic Development and Injury Response J. Neurosci., 2018-02-26;0(0):. 2018-02-26 [PMID: 29483282] (Bioassay, Mouse)
• 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 The inhibitory effect of IFN-gamma on protease HTRA1 expression in rheumatoid arthritis. J Immunol, 2014-06-06;193(1):130-8. 2014-06-06 [PMID: 24907345] (ELISA (Standard))

Bioinformatics

• Gene Symbol: HTRA1
• Uniprot:
  • Q92743()