|ABCA1 in HepG2 Human Cell Line. ABCA1 was detected in immersion fixed HepG2 human hepatocellular carcinoma cell line using Sheep Anti-Human ABCA1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF7207) at 10 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Sheep IgG Secondary Antibody (red, upper panel; Catalog # NL010) and counterstained with DAPI (blue, lower panel). View our protocol for Fluorescent ICC Staining of Cells on Coverslips.|
Intracellular Staining by Flow Cytometry
Detection of ABCA1 in HepG2 Human Cell Line by Flow Cytometry. |
HepG2 human hepatocellular carcinoma cell line was stained with Sheep Anti-Human ABCA1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF7207, filled histogram) or control antibody (Catalog # 5-001-A, open histogram), followed by Allophycocyanin-conjugated Anti-Sheep IgG Secondary Antibody (Catalog # F0127). To facilitate intracellular staining, cells were fixed with paraformaldehyde and permeabilized with saponin.
ABCA1 (ATP-binding cassette, subfamily A member 1; also cholesterol efflux regulatory protein) is a 240-280 kDa member of the ABCA family, ABC transporter superfamily of molecules. It is widely expressed, particularly by macrophages, where it participates in the transport of lipid across cell membranes. Once transported, it is passed on to tethered apoA-I to form HDL. Human ABCA1 is 2261 amino acids (aa) in length. It has up to 15 transmembrane segments with beginning and ending cytoplasmic domains and a large cytoplasmic central region. ABC transporters are found between aa 899-1311 and 1912-2144. Amino acids 2216-2221 are essential for lipid transport, and two intramolecular disulfide bonds are required for activity (Cys75-Cys309 and Cys1463/1465-Cys1477). There are three potential isoform variants. One possesses an alternative start site at Met61, while two others contain 12 aa and 123 aa substitutions for aa 241-2261, respectively. Over aa 1248-1350, human ABCA1 shares 97% aa identity with mouse ABCA1.