Recombinant Anti-TFEB antibody [BLR070G] (ab264421)

False colour image of Western blot: Anti-TFEB antibody [BLR070G] 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, ab264421 was shown to bind specifically to TFEB. A band was observed at 60 kDa in wild-type HepG2 cell lysates with no signal observed at this size in TFEB knockout cell line ab277871 (knockout cell lysate ab284220). To generate this image, wild-type and TFEB knockout HepG2 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.

Lysates prepared using NETN lysis buffer.

TFEB was immunoprecipitated from HeLa (human epithelial cell line from cervix adenocarcinoma) whole cell lysate (1.0 mg per IP reaction; 20% of IP loaded) with ab264421 at 20 µl/mg lysate. Western blot was performed from the immunoprecipitates using ab264421 at 1/1000 dilution.

Lane 1: ab264421 IP in HeLa whole cell lysate.
Lane 2: Control IgG IP in HeLa whole cell lysate.

Detection: Chemiluminescence with an exposure time of 75 seconds.

Lysates prepared using NETN lysis buffer.