Recombinant Human Biotinidase/BTD Protein, CF

R&D Systems | Catalog # 7839-BT

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Key Product Details

  • R&D Systems CHO-derived Recombinant Human Biotinidase/BTD Protein (7839-BT)
  • 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

CHO

Accession Number

P43251

Applications

Enzyme Activity
View all Biotinidase/BTD Proteins and Enzymes »

Product Specifications

Source

Chinese Hamster Ovary cell line, CHO-derived human Biotinidase/BTD protein
Ala42-Asp543, with a C-terminal 6-His tag

Purity

>95%, by SDS-PAGE under reducing conditions and visualized by Colloidal Coomassie® Blue stain at 5 μg per lane.

Endotoxin Level

<1.0 EU per 1 μg of the protein by the LAL method.

N-terminal Sequence Analysis

Ala42

Predicted Molecular Mass

58 kDa

SDS-PAGE

65-80 kDa, reducing conditions

Activity

Measured by its ability to hydrolyze the substrate biotin 4-Nitrophenyl ester (BNP).
The specific activity is >190 pmol/min/μg, as measured under the described conditions.

Formulation, Preparation, and Storage

7839-BT
Formulation Supplied as a 0.2 μm filtered solution in NaH2PO4, NaCl, EDTA 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: Biotinidase/BTD

Biotinidase (BTD) is a member of the nitrilase superfamily, which consists of 12 families of nitrilases, amidases, carbamylases, and N‑acyltrasferases (1). It is a thiol hydrolase releasing biotin from biotinamide, biotin‑lysine, biotin‑peptide conjugates and biotin methylester. It is expressed in most mammalian tissues, with high activity being present in liver, kidney, serum, intestine, and adrenal glands (2). BTD with two other proteins, sodium-dependent multivitamin transporter, and holocarboxylase synthetase, play major roles in the homeostasis of biotin (3). BTD contributes to the homeostasis through the intestinal release of free biotin from digested biotin‑containing proteins and plasma transport and the recycling of biotin from breakdown products of biotinylated carboxylases. BTD deficiency can lead to a decrease in biotin bioavailability due to failure in releasing biotin from dietary proteins. It can be caused by gene mutations or from decreased secretion of BTD into the intestinal lumen (4). A recent study indicates that the BTD level in human plasma is a potential biomarker for the detection of breast cancer (5).

References

  1. Pace, H. C. and C. Brenner (2001) Genome. Biol. 2:reviews/0001.1.
  2. Chauhan, J. and K. Dakshinamurt (1986) J. Biol. Chem. 261:4286.
  3. Wolf, B. (2005) J Nur Biochem. 16:441.
  4. Zempleni, J. et al. (2008) Expert Rev Endocrinol Metab. 3:715.
  5. Kang, U. B. et al. (2010) BMC Cancer 10:114.

Alternate Names

Biotinase, BTD

Entrez Gene IDs

686 (Human); 26363 (Mouse); 306262 (Rat)

Gene Symbol

BTD

UniProt

P43251 (Human)

Additional Biotinidase/BTD Products

Product Documents for Recombinant Human Biotinidase/BTD 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 Biotinidase/BTD Protein, CF

For research use only

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Protocols

View specific protocols for Recombinant Human Biotinidase/BTD Protein, CF (7839-BT):

Materials
  • Assay Buffer: 50 mM Tris, 0.2 M NaCl, 0.1% Triton, pH 7.5
  • Recombinant Human Biotinidase/BTD (rhBTD) (Catalog # 7839-BT)
  • (+)-Biotin 4-Nitrophenyl ester (BNP) (Sigma, Catalog # 861650), 50 mM stock in DMSO
  • Dimethyl Sulfoxide (DMSO) (Sigma, Catalog # 34869)
  • 96-well Clear Plate (Catalog # DY990)
  • Plate Reader (Model: SpectraMax Plus by Molecular Devices) or equivalent
  1. Dilute rhBTD to 20 µg/mL in Assay Buffer.
  2. Dilute room temperature BNP to 1 mM in DMSO. Mix well.
  3. Load 50 µL of 20 µg/mL rhBTD in a clear strip well plate, and start the reaction by adding 50 µL of 1 mM BNP. Include a Substrate Blank containing 50 µL Assay Buffer and 50 µL of 1 mM BNP.
  4. Incubate sealed plate at room temperature for 10 minutes in the dark.
  5. Read at 405 nm (absorbance) in endpoint mode.
  6. Calculate specific activity:

Specific Activity (pmol/min/µg) =

Adjusted Abs* (OD) x Conversion Factor** (pmol/OD)
Incubation time (min) x amount of enzyme (µg)

*Adjusted for Substrate Blank

**Derived using calibration standard p-Nitrophenol (Sigma, Catalog # 241326). Per Well:

  • rhBTD: 1 µg
  • BNP: 0.5 mM

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