Recombinant Human Biliverdin Reductase B/BLVRB Protein, CF

R&D Systems | Catalog # 6568-BR

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Key Product Details

  • R&D Systems E. coli-derived Recombinant Human Biliverdin Reductase B/BLVRB Protein (6568-BR)
  • 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

P30043

Applications

Enzyme Activity
View all Biliverdin Reductase B/BLVRB Proteins and Enzymes »

Product Specifications

Source

E. coli-derived human Biliverdin Reductase B/BLVRB protein
Ala2-Gln206, 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

N-terminus confirmed by detection of His tag using Western analysis.

Predicted Molecular Mass

23 kDa

SDS-PAGE

23-25 kDa, reducing conditions

Activity

Measured by the reduction of riboflavin 5'-monophosphate (FMN) using NADPH as the cofactor.
The specific activity is >225 pmol/min/μg, as measured under the described conditions.

Formulation, Preparation, and Storage

6568-BR
Formulation Supplied as a 0.2 μm filtered solution in Tris, NaCl, Brij and Glycerol.
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.

Background: Biliverdin Reductase B/BLVRB

Clearance of heme in mammals is a two-step process starting with conversion of heme to biliverdin by heme oxygenase, followed by reduction of biliverdin to bilirubin by bilivredin reductase. Biliverdin IX b reductase (BLVRB) converts the  beta  isomer of biliverdin IX to bilirubin IX b, which constitutes 87% of total bilirubin in fetal bile. Therefore BLVRB is especially important for fetal heme metabolism and clearance (1). BLVRB is a cytoplasmic enzyme expressed at high levels in erythrocytes and liver, but is present in other tissues (2). The enzyme is identical to flavin reductase, which is an oxidoreductase that catalyses the NADPH-dependent reduction for a variety of flavins, such as riboflavin, FAD or FMN and met‑hemoglobin (3, 4). BLVRB is structurally distinct from BLVRA. In contrast to BLVRA, which prefers the biliverdin alpha  isomer but could also use the  beta  isomer as substrate, BLVRB is specific for the beta  isomer (5, 6).

References

  1. Pereira, P.J. et al. (2001) Nat. Struct. Biol. 8:215.
  2. Chikuba, K. et al. (1994) Biochem. Biophys. Res. Commun. 198:1170.
  3. Shalloe, F. et al. (1996) Biochem. J. 316:385.
  4. Cunningham, O. et al. (2000) Biochem. J. 345:393.
  5. Yamaguchi, T. et al. (1994) J. Biol. Chem. 269:24343.
  6. Cunningham, O. et al. (2000) J. Biol. Chem. 275:19009.

Alternate Names

BLVRB, BVRB, FLR, GHBP

Entrez Gene IDs

645 (Human)

Gene Symbol

BLVRB

UniProt

P30043

Additional Biliverdin Reductase B/BLVRB Products

Product Documents for Recombinant Human Biliverdin Reductase B/BLVRB 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 Human Biliverdin Reductase B/BLVRB Protein, CF

For research use only

Related Research Areas

Redox Enzymes

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Protocols

View specific protocols for Recombinant Human Biliverdin Reductase B/BLVRB Protein, CF (6568-BR):

Materials
  • Assay Buffer: 100 mM Sodium Acetate, pH 5.0
  • Recombinant Human Biliverdin Reductase B/BLVRB (rhBLVRB) (Catalog # 6568-BR)
  • Riboflavin 5’-monophosphate sodium salt dihydrate (FMN) (Sigma, Catalog # F6750), 10 mM in deionized water
  • beta -Nicotinamide adenine dinucleotide phosphate reduced, tetrasodium salt ( beta -NADPH) (Sigma, Catalog # N7505), 10 mM in deionized water
  • UV Plate (Costar, Catalog # 3635)
  • Plate Reader (Model: SpectraMax Plus by Molecular Devices) or equivalent
  1. Dilute rhBLVRB to 40 ng/μL in Assay Buffer.
  2. Prepare a Reaction Mixture by combining FMN and beta -NADPH in Assay Buffer to a concentration of 400 μM for each.
  3. Load 50 μL of 40 ng/μL rhBLVRB into the microplate, and start the reaction by adding 50 μL of Reaction Mixture. Include a Substrate Blank containing 50 μL of Assay Buffer and 50 μL of Reaction Mixture.
  4. Read at an absorbance of 340 nm in kinetic mode for 5 minutes (Note: readings will be negative).
  5. Calculate specific activity:

     Specific Activity (pmol/min/µg) =

 Adjusted Vmax* (OD/min) x -1 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 6270 M-1cm-1 
     ***Using the path correction 0.32 cm
     Note: the output of many spectrophotometers is in mOD. Per Well:
  • rhBLVRB: 2 μg
  • FMN: 200 μM
  • beta -NADPH: 200 μM

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