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Recombinant Human HO-1/HMOX1 Protein, CF

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Product Details

Summary
Reactivity HuSpecies Glossary
Applications Enzyme Activity
Format
Carrier-Free

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Recombinant Human HO-1/HMOX1 Protein, CF Summary

Details of Functionality
Measured by its ability to oxidize hemin to biliverdin. The specific activity is >5 pmol/min/μg, as measured under the described conditions.
Source
E. coli-derived human HO-1/HMOX1/HSP32 protein
Met1-Thr261 (Phe33Leu), with a C-terminal 6-His tag
Accession #
N-terminal Sequence
Met1
Protein/Peptide Type
Recombinant Enzymes
Gene
HMOX1
Purity
>95%, 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
31 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
29-31 kDa, reducing conditions
Publications
Read Publication using
3776-HM in the following applications:

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 Tris, NaCl and Glycerol.
Purity
>95%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.
Assay Procedure
  • Assay Buffer: 50 mM Tris, pH 7.5
  • Recombinant Human HO‑1/HMOX1/HSP32 (rhHO-1) (Catalog # 3776-HM)
  • Recombinant Human POR/Cytochrome P450 Reductase (rhPOR) (Catalog # 6340-PR)
  • Recombinant Human Biliverdin Reductase A/BLVRA (rhBLVRA) (Catalog # 6454-BR)
  • Catalase (Sigma, Catalog # C30)
  • Hemin (Sigma, Catalog # H9039), 10 mM stock in DMSO
  • Bovine Serum Albumin (BSA), 100 mg/mL stock in deionized water
  • beta -Nicotinamide adenine 2’-phosphate reduced tetrasodium salt hydrate (NADPH) (Sigma, Catalog # N7505), 10 mM stock in deionized water
  • 96-well Clear Plate (Costar, Catalog # 92592)
  • Plate Reader (Model: SpectraMax Plus by Molecular Devices) or equivalent
  1. Dilute Hemin to 1 mM in Assay Buffer.
  2. Prepare Reaction Mixture containing 60 µM Hemin, 4 mg/mL BSA, 1000 U/mL catalase, 80 µg/mL rhPOR, and 80 µg/mL rhBLVRA in Assay Buffer.
  3. Prepare 80 µg/mL rhHO-1 in Assay Buffer.
  4. Dilute NADPH to 1 mM in Assay Buffer.
  5. In a clear plate, load 25 µL dilute rhHO-1 and add 25 µL Reaction Mixture. Include an enzyme blank containing 25 µL Assay Buffer and 25 µL Reaction Mixture.
  6. Start the reaction by adding 50 µL 1 mM NADPH to all wells used.
  7. Read absorbance at 468 nm (bottom read) in kinetic mode for 5 minutes.
  8. Calculate specific activity:

     Specific Activity (pmol/min/µg) =

 Adjusted Vmax* (OD/min) x well volume (L) x 1012 pmol/mol
ext. coeff** (M-1cm-1) x path corr.*** (cm) x amount of enzyme (µg)

     *Adjusted for Substrate Blank 
     **Using the extinction coefficient 43500 M-1cm-1 
     ***Using the path correction 0.32 cm
     Note: the output of many spectrophotometers is in mOD Per Reaction:
  • rhHO-1: 2 µg
  • rhPOR: 2 µg
  • rhBLVRA: 2 µg
  • Catalase: 250 U/mL
  • Hemin: 15 µM
  • BSA: 1 mg/mL
  • NADPH: 0.5 mM

Notes

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

Alternate Names for Recombinant Human HO-1/HMOX1 Protein, CF

  • bK286B10
  • EC 1.14.99.3
  • heat shock protein, 32-kD
  • heme oxygenase (decycling) 1
  • HMOX1
  • HO
  • HO1
  • HO-1
  • HO-1heme oxygenase 1
  • HSP32

Background

Heme oxygenase (HMOX) is the rate limiting enzyme in heme catabolism (1). It cleaves heme to biliverdin, carbon monoxide, and iron. The biliverdin is subsequently converted to bilirubin by biliverdin reductase. The mechanism of HMOX is unique in that heme serves as the substrate of the enzyme and as the prosthetic group for the activation of iron-bound O2. HMOX activity is highest in spleen where senescent erythrocytes are sequestered and destroyed. Two isoforms, HMOX1 and HMOX2, are expressed in most tissues. HMOX1 is an inducible enzyme in response to heme, heavy metals, oxidative stress, cytokines, and many drugs (2). Whereas HMOX2 displays a constitutive expression. HMOX1 is expressed mainly in spleen, liver, and kidney, and HMOX2 is prominently expressed in the brain and testes. The increased expression of HMOX1 levels is related to a variety of pathological states, where it functions as a cytoprotective molecule through its by‑products (3). HMOX1 also plays important roles in the regulation of cell proliferation, differentiation, and apoptosis.
  1. Kappas, A. (2008) Pharmacol. Rev. 60:79.
  2. Otterbein, L.E. and Choi, A.M.K. (2000) Am. J. Physiol. Lung Cell. Mol. Physiol. 279:1029.
  3. Grochot-Przeczek, A. et al. (2012) Clin. Sci. (Lond) 122:93.

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Publications for HO-1/HMOX1/HSP32 (3776-HM)(1)

We have publications tested in 1 confirmed species: N/A.

We have publications tested in 1 application: Bioassay.


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Bioinformatics

Gene Symbol HMOX1
Uniprot