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Recombinant Mouse RBP4 Protein, CF

Catalog #: 3476-LC
2 Citations Datasheet / COA / SDS
Catalog # Availability Size / Price Qty
3476-LC-050
R&D Systems Recombinant Proteins and Enzymes
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Citations (2)
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Summary Product Datasheets Carrier Free Data Images Reconstitution Calculator Background Related Research Areas

Recombinant Mouse RBP4 Protein, CF Summary

Product Specifications

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.
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.
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 #
Q00724
N-terminal Sequence
Analysis
Glu19
Predicted Molecular Mass
22 kDa
SDS-PAGE
22 kDa, reducing conditions

Product Datasheets

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3476-LC

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.

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.

Assay Procedure

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
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: 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.
Entrez Gene IDs
5950 (Human); 19662 (Mouse); 25703 (Rat)
Alternate Names
interstitial; Plasma retinol-binding protein; RBP4; retinol binding protein 4, plasma; RetinolBinding Protein 4; retinol-binding protein 4, plasma

Citations for Recombinant Mouse RBP4 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.

2 Citations: Showing 1 - 2
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  1. Alteration of circadian machinery in monocytes underlies chronic kidney disease-associated cardiac inflammation and fibrosis
    Authors: Y Yoshida, N Matsunaga, T Nakao, K Hamamura, H Kondo, T Ide, H Tsutsui, A Tsuruta, M Kurogi, M Nakaya, H Kurose, S Koyanagi, S Ohdo
    Nature Communications, 2021-05-13;12(1):2783.
    Species: Mouse
    Sample Types: Cell Lysates
    Applications: Reference Standard
  2. Adipose tissue exosome-like vesicles mediate activation of macrophage-induced insulin resistance.
    Authors: Deng ZB, Poliakov A, Hardy RW, Clements R, Liu C, Liu Y, Wang J, Xiang X, Zhang S, Zhuang X, Shah SV, Sun D, Michalek S, Grizzle WE, Garvey T, Mobley J, Zhang HG
    Diabetes, 2009-08-12;58(11):2498-505.
    Species: Mouse
    Sample Types: Exosomes
    Applications: Bioassay

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