Recombinant Mouse RBP4 Protein, CF Summary
Glu19-Leu201, with a C-terminal Ile and 6-His tag
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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.
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.
|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.
- 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
- Dilute rmRBP4 to 49 µg/mL (2.2 µM) in Assay Buffer.
- Make serial dilutions of retinoic acid in 95% ethanol to 100, 30, 10, 3 and 1 µM.
- The formation of protein/retinol binding in microtubes:
- Mix 112.5 µL of diluted rmRBP4 and 12.5 µL of retinoic acid serial dilutions in microtubes.
- For a blank, mix 112.5 µL rmRBP4 and 12.5 µL of 95% ethanol in a microtube.
- Incubate at room temperature for 30 minutes.
- Load into a black well plate 100 µL of the reaction mixtures and blank.
- Read at excitation and emission wavelengths of 280 nm and 340 nm (top read), respectively, in endpoint mode.
- Calculate the 50% binding concentration (BC50) for retinoic acid by plotting RFU vs. concentration with 4‑PL fitting.
- rmRBP4: 4.41 µg
- Retinoic Acid Curve: 10, 3, 1, 0.3 and 0.1 µM
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.
- Zanotti, G. and R. Berni (2004) Vitamins and Hormones 69:271.
- Newcomer, M.E. and D.E. Ong (2000) Biochim. Biophys. Acta 1482:57.
- Yang, Q. et al. (2005) Nature 436:356.
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.
Citations: Showing 1 - 2
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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;12(1):2783.
Sample Types: Cell Lysates
Applications: Reference Standard
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
Sample Types: Exosomes
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