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Recombinant Mouse RBP4 Protein, CF

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Summary
Reactivity MuSpecies Glossary
Applications Binding Activity
Format
Carrier-Free

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Recombinant Mouse RBP4 Protein, CF Summary

Details of Functionality
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.
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
Accession #
N-terminal Sequence
Glu19
Protein/Peptide Type
Recombinant Enzymes
Gene
Rbp4
Purity
>95%, by SDS-PAGE under reducing conditions and visualized by silver stain
Endotoxin Note
<1.0 EU per 1 μg of the protein by the LAL method.

Applications/Dilutions

Dilutions
  • Binding Activity
Theoretical MW
22 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
22 kDa, reducing conditions
Publications
Read Publications using
3476-LC 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 reconstitution.
Buffer
Lyophilized from a 0.2 μm filtered solution in Tris and NaCl.
Purity
>95%, by SDS-PAGE under reducing conditions and visualized by silver stain
Reconstitution Instructions
Reconstitute at 500 μg/mL in sterile 50 mM Tris, 10 mM CaCl2 and 150 mM NaCl, pH 7.5.
Assay Procedure
  • 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

Notes

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

Alternate Names for Recombinant Mouse RBP4 Protein, CF

  • interstitial
  • Plasma retinol-binding protein
  • RBP4
  • retinol binding protein 4, plasma
  • RetinolBinding Protein 4
  • retinol-binding protein 4, plasma

Background

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.

  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.

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Publications for RBP4/Retinol-Binding Protein 4 (3476-LC)(3)

We have publications tested in 2 confirmed species: Mouse, Transgenic Mouse.

We have publications tested in 2 applications: Bioassay, Reference Standard.


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(2)
Reference Standard
(1)
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(3)
Transgenic Mouse
(1)
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Bioinformatics

Gene Symbol Rbp4
Uniprot