Intracellular Staining by Flow Cytometry
|Detection of RBP4/Retinol-Binding Protein 4 in HepG2 Human Cell Line by Flow Cytometry. HepG2 human hepatocellular carcinoma cell line was stained with Mouse Anti-Human RBP4/Retinol‑Binding Protein 4 Monoclonal Antibody (Catalog # MAB3378, filled histogram) or isotype control antibody (Catalog # MAB002, open histogram), followed by Phycoerythrin-conjugated Anti-Mouse IgG F(ab')2 Secondary Antibody (Catalog # F0102B). To facilitate intracellular staining, cells were fixed with paraformaldehyde and permeabilized with methanol.|
Retinol (also known as vitamin A) is unstable and insoluble in the aqueous solution (1, 2). 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. 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.
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