Recombinant Anti-LIMPII antibody [EPR12080] (ab176317)

False colour image of Western blot: Anti-LIMPII antibody [EPR12080] staining at 1/1000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab176317 was shown to bind specifically to LIMPII. A band was observed at 80 kDa in wild-type MCF7 cell lysates with no signal observed at this size in SCARB2 knockout cell line ab274952 (knockout cell lysate ab275010). To generate this image, wild-type and SCARB2 knockout MCF7 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) at 1/20000 dilution.

Western blot analysis on immunoprecipitation pellet from Y79 (Human retinoblastoma cell line) cell lysate labeling LIMPII using ab176317 at a 1/10 dilution.

Immunohistochemical analysis of paraffin-embedded human kidney tissue labling LIMPII using ab176317 at a 1/50 dilution.

Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

Immunohistochemical analysis of paraffin-embedded human liver tissue labling LIMPII using ab176317 at a 1/50 dilution.

Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

Intracellular flow cytometric analysis of permeabilized HeLa cells labeling LIMPII using ab176317 at a 1/10 dilution (red)or a rabbit IgG control (green).