Recombinant Human HTRA1 Protein, CF

Catalog # Availability Size / Price Qty
2916-SE-020
R&D Systems Recombinant Proteins and Enzymes
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Product Details
Citations (5)
FAQs
Reviews (1)

Recombinant Human HTRA1 Protein, CF Summary

Product Specifications

Purity
>90%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.
Endotoxin Level
<1.0 EU per 1 μg of the protein by the LAL method.
Activity
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 #
N-terminal Sequence
Analysis
Met
Predicted Molecular Mass
36 kDa
SDS-PAGE
37-40 kDa, reducing conditions

Product Datasheets

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2916-SE

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.

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.
  • 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 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
Reconstitution Calculator

Reconstitution Calculator

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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.

References
  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.
Long Name
High Temperature Requirement Protein-1
Entrez Gene IDs
5654 (Human)
Alternate Names
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)

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.

5 Citations: Showing 1 - 5
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  1. 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;11(8):674.
    Species: Human
    Sample Types: Protein
    Applications: Bioassay
  2. 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;11(8):674.
    Species: Human
    Sample Types: Protein
    Applications: Bioassay
  3. 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;19(5):.
    Species: Human
    Sample Types: Whole Cells
    Applications: Bioassay
  4. 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;0(0):.
    Species: Mouse
    Sample Types: Whole Cells
    Applications: Bioassay
  5. 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;193(1):130-8.
    Species: N/A
    Sample Types: N/A
    Applications: ELISA (Standard)

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Recombinant Human HTRA1 Protein, CF
By Anonymous on 09/19/2018
Application: In vitro bioactivity in cell culture