Recombinant Mouse RBP4 Protein, CF

R&D Systems | Catalog # 3476-LC

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Key Product Details

  • R&D Systems NS0-derived Recombinant Mouse RBP4 Protein (3476-LC)
  • 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

NS0

Accession Number

Q00724

Applications

Binding Activity
View all RBP4/Retinol-Binding Protein 4 Proteins and Enzymes »

Product Specifications

Source

Mouse myeloma cell line, NS0-derived mouse RBP4/Retinol-Binding Protein 4 protein
Glu19-Leu201, with a C-terminal Ile and 6-His tag

Purity

>95%, by SDS-PAGE under reducing conditions and visualized by silver stain.

Endotoxin Level

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

N-terminal Sequence Analysis

Glu19

Predicted Molecular Mass

22 kDa

SDS-PAGE

22 kDa, reducing conditions

Activity

Measured by its ability to bind all-trans retinoic acid. The binding of retinoic acid results in the quenching of Trp fluorescence in RBP4.
The 50% binding concentration (BC50) is >0.5 µM, as measured under the described conditions.

Formulation, Preparation, and Storage

3476-LC
Formulation Lyophilized from a 0.2 μm filtered solution in Tris and NaCl.
Reconstitution Reconstitute at 500 μg/mL in sterile 50 mM Tris, 10 mM CaCl2 and 150 mM NaCl, pH 7.5.
Shipping The product is shipped at ambient temperature. 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 reconstitution.

Calculators

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: RBP4/Retinol-Binding Protein 4

Retinol (also known as vitamin A) is unstable and insoluble in the aqueous solution. However, retinol becomes quite stable and soluble in plasma due to its tight interaction with retinol‑binding protein 4 (RBP4), also known as plasma retinol‑binding protein (1-3). A prototypic member of the lipocalin superfamily, RBP4 has a
beta ‑barrel structure with a well-defined cavity. It is secreted from the liver, a process requiring the availability of retinol. RBP4 delivers retinol from the liver to the peripheral tissues. In plasma, the RBP4‑retinol complex interacts with transthyretin (TTR), also known as thyroxine-binding protein and prealbumin. The retinol‑RBP4‑TTR complex prevents the loss of RBP4 by filtration through the kidney and increases the stability of the retinol‑RBP4 complex. Defects in RBP4 cause retinol‑binding protein deficiency, which affects night vision. Serum RBP4 levels are elevated in insulin-resistant mice and humans with obesity and type 2 diabetes, implying that RBP4, an adipocyte-derived signal, may be a biomarker and a drug target for the two diseases.

References

  1. Zanotti, G. and R. Berni (2004) Vitamins and Hormones 69:271.
  2. Newcomer, M.E. and D.E. Ong (2000) Biochim. Biophys. Acta 1482:57.
  3. Yang, Q. et al. (2005) Nature 436:356.

Alternate Names

RetinolBinding Protein 4

Entrez Gene IDs

5950 (Human); 19662 (Mouse); 25703 (Rat)

Gene Symbol

RBP4

UniProt

Q00724

Additional RBP4/Retinol-Binding Protein 4 Products

Product Documents for Recombinant Mouse RBP4 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 RBP4 Protein, CF

For research use only

Citations for Recombinant Mouse RBP4 Protein, CF

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Protocols

View specific protocols for Recombinant Mouse RBP4 Protein, CF (3476-LC):

Materials
  • Assay Buffer: 50 mM Tris, 10 mM CaCl2, 150 mM NaCl, pH 7.5 (TCN)
  • Recombinant Mouse RBP4/Retinol‑Binding Protein 4 (rmRBP4) (Catalog # 3476-LC)
  • 95% Ethanol
  • Retinoic Acid (Tocris, Catalog # 0695), 50 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 rmRBP4 to 49 µg/mL (2.2 µM) in Assay Buffer.
  2. Make serial dilutions of retinoic acid in 95% ethanol to 100, 30, 10, 3 and 1 µM.
  3. The formation of protein/retinol binding in microtubes:
    1. Mix 112.5 µL of diluted rmRBP4 and 12.5 µL of retinoic acid serial dilutions in microtubes.
    2. For a blank, mix 112.5 µL rmRBP4 and 12.5 µL of 95% ethanol in a microtube.
    3. Incubate at room temperature for 30 minutes.
  4. Load into a black well plate 100 µL of the reaction mixtures and blank.
  5. Read at excitation and emission wavelengths of 280 nm and 340 nm (top read), respectively, in endpoint mode.
  6. Calculate the 50% binding concentration (BC50) for retinoic acid by plotting RFU vs. concentration with 4‑PL fitting.

Per Well:

  • rmRBP4: 4.41 µg
  • Retinoic Acid Curve: 10, 3, 1, 0.3 and 0.1 µM

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