Recombinant Mouse BLMH/Bleomycin Hydrolase Protein, CF

R&D Systems | Catalog # 6180-CY

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Key Product Details

  • R&D Systems E. coli-derived Recombinant Mouse BLMH/Bleomycin Hydrolase Protein (6180-CY)
  • Quality control testing to verify active proteins with lot specific assays by in-house scientists
  • All R&D Systems proteins are covered with a 100% guarantee

Source

E. coli

Accession Number

Q8R016

Applications

Enzyme Activity
View all BLMH/Bleomycin Hydrolase Proteins and Enzymes »

Product Specifications

Source

E. coli-derived mouse BLMH/Bleomycin Hydrolase protein
Asn2-Glu455 with an N-terminal Met and 6-His tag

Purity

>95%, 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.

N-terminal Sequence Analysis

Met

Predicted Molecular Mass

53 kDa

SDS-PAGE

45-55 kDa, reducing conditions

Activity

Measured by its ability to hydrolyze Met-AMC.
The specific activity is >1,500 pmol/min/μg, as measured under the described conditions.

Formulation, Preparation, and Storage

6180-CY
Formulation Supplied as a 0.2 μm filtered solution in Tris, NaCl, Glycerol and DTT.
Shipping The product is shipped with dry ice or equivalent. 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, -70 °C as supplied.
  • 3 months, -70 °C under sterile conditions after opening.

Background: BLMH/Bleomycin Hydrolase

Bleomycin hydrolase (BLMH) is a cysteine peptidase of the papain superfamily. It is named for its ability to hydrolyze the anti‑tumor agent bleomycin and inactivate it (1). It has a papain-like catalytic triad (Cys-His-Asp) with optimum activity at neutral pH. In mammals it is expressed ubiquitously in all types of tissues and its expression is up‑regulated in many tumors. It is present in the cytoplasm as homohexameric protein of approximately 300 kDa. In addition to its aminopeptidase activity, it has homocysteine-thiolactonase activity. The normal physiological function of BLMH is not clear. BLMH inactivates bleomycin, a glycopeptide anti‑cancer agent, by deaminating it (2). BLMH has been suggested to play a role in the generation of MHC class I-presented peptides (3, 4). Diminished BLMH activity may contribute to the pathology of Alzheimer’s disease (AD) (5, 6). It is inhibited by cysteine protease inhibitors such as N‑ethylmaleimide, iodoacetamide, para‑hydroxymercuribenzoate, and E-64.

References

  1. Joshua-Tor, L. and S. A. Johnson (2004) in Handbook of Proteolytic Enzymes, Barrett, A. J. et al. eds. pp. 1197.
  2. Schwartz, D. R. et al. (1999) Proc. Natl. Acad. Sci. USA, 96:4680.
  3. Kim, E. et al. (2009) J. Immunol. 183:7379.
  4. Towne, C. F. et al. (2007) J. Immunol. 178:6923.
  5. Suszynska, J. et al. (2010) J. Alzheimers Dis. 19:1177.
  6. Lefterov, I. M. et al. (2000) FASEB J. 14:1837.

Alternate Names

BMH

Entrez Gene IDs

642 (Human)

Gene Symbol

BLMH

UniProt

Q8R016

Additional BLMH/Bleomycin Hydrolase Products

Product Documents for Recombinant Mouse BLMH/Bleomycin Hydrolase Protein, CF

Certificate of Analysis

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Note: Certificate of Analysis not available for kit components.

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Product Specific Notices for Recombinant Mouse BLMH/Bleomycin Hydrolase Protein, CF

Coomassie is a registered trademark of Imperial Chemical Industries Ltd.

For research use only

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Protocols

View specific protocols for Recombinant Mouse BLMH/Bleomycin Hydrolase Protein, CF (6180-CY):

Materials
  • Assay Buffer: 50 mM HEPES, 5 mM EDTA, 10 mM DTT, pH 7.0
  • Recombinant Mouse BLMH/Bleomycin Hydrolase  (rmBLMH) (Catalog # 6180-CY)
  • Substrate: Met-AMC (Bachem, Catalog # I-1265), 100 mM stock in DMSO
  • F16 Black Maxisorp Plate (Nunc, Catalog # 475515)
  • Fluorescent Plate Reader (Model: SpectraMax Gemini EM by Molecular Devices) or equivalent
  1. Dilute rmBLMH to 10 µg/mL in Assay Buffer.
  2. Incubate at 37 °C for 30 minutes.
  3. Dilute Activated rmBLMH to 0.4 µg/mL.
  4. Dilute Substrate to 2 mM in Assay Buffer.
  5. Load 50 µL of the 0.4 µg/mL rmBLMH into a plate, and start the reaction by adding 50 µL of 2 mM Substrate. Include a Substrate Blank containing 50 µL Assay Buffer and 50 µL of 2 mM Substrate without any rmBLMH.
  6. Read at excitation and emission wavelengths of 380 nm and 460 nm (top read), respectively, in kinetic mode for 5 minutes.
  7. Calculate specific activity:

     Specific Activity (pmol/min/µg) =

 Adjusted Vmax* (RFU/min) x Conversion Factor** (pmol/RFU)
amount of enzyme (µg)

     *Adjusted for Substrate Blank
     **Derived using calibration standard 7-Amino, 4-Methyl Coumarin (AMC) (Sigma, Catalog # A-9891).

Per Well:
  • rmBLMH: 0.02 µg
  • Substrate: 1 mM

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