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Recombinant Human Biliverdin Reductase B/BLVRB Protein, CF

Catalog #: 6568-BR
1 Citation Datasheet / COA / SDS
Catalog # Availability Size / Price Qty
6568-BR-020
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Summary Product Datasheets Carrier Free Data Images Reconstitution Calculator Background Related Research Areas

Recombinant Human Biliverdin Reductase B/BLVRB Protein, CF Summary

Product Specifications

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.
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.
Source
E. coli-derived human Biliverdin Reductase B/BLVRB protein
Ala2-Gln206, with an N-terminal Met and 6-His tag
Accession #
P30043
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

Product Datasheets

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6568-BR

Product Datasheet
COA
SDS

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.

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.

Assay Procedure

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
Reconstitution Calculator

Reconstitution Calculator

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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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.
Entrez Gene IDs
645 (Human)
Alternate Names
biliverdin reductase B (flavin reductase (NADPH)); Biliverdin Reductase B; Biliverdin-IX beta-reductase; BLVRB; BVRB; BVR-B; EC 1.3.1.24; EC 1.5.1.30; flavin reductase (NADPH); flavin reductase; FLR; FLRBVRB; FR; GHBP; Green heme-binding protein; MGC117413; NADPH-dependent diaphorase; NADPH-flavin reductase; SDR43U1; short chain dehydrogenase/reductase family 43U, member 1

Citation for Recombinant Human Biliverdin Reductase B/BLVRB 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.

1 Citation: Showing 1 - 1

  1. Correlating chemical sensitivity and basal gene expression reveals mechanism of action.
    Authors: Rees M, Seashore-Ludlow B, Cheah J, Adams D, Price E, Gill S, Javaid S, Coletti M, Jones V, Bodycombe N, Soule C, Alexander B, Li A, Montgomery P, Kotz J, Hon C, Munoz B, Liefeld T, Dancik V, Haber D, Clish C, Bittker J, Palmer M, Wagner B, Clemons P, Shamji A, Schreiber S
    Nat Chem Biol, 2015-12-14;12(2):109-16.
    Species: Human
    Sample Types: Recombinant Protein
    Applications: Bioassay

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