Anti-LRP6 antibody [EPR22910-39]

• WB

Lab

Western blot - Anti-LRP6 antibody [EPR22910-39] (AB231779)

The molecular weight observed is consistent with what has been described in the literature (PMID : 16989816, 16263759, 28341812). ab231779 was shown to specifically react with LRP6 in wild-type HAP1 cells as signal was lost in LRP6 knockout cells. Wild-type and LRP6 knockout samples were subjected to SDS-PAGE. ab231779 and ab129002 (Rabbit anti-Vinculin loading control) were incubated 1 hour at room temperature at 1/1000 dilution and 1/200,000 dilution respectively. Blots were developed with Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) secondary antibody at 1/20,000 dilution for 1 hour at room temperature before imaging. The blot was developed on a BIO-RAD^® ChemiDoc™ instrument using the ECL technique. This blot was developed using a higher sensitivity ECL substrate.

Blocking/Dilution buffer : 5% NFDM/TBST.

Exposure times : Lanes 1-2 : 3 mins; Lanes 3 : 26 secs; Lanes 4-5 : 3 mins.

All lanes:

Western blot - Anti-LRP6 antibody [EPR22910-39] (ab231779) at 1/1000 dilution

Lane 1:

Wild-type HAP1 whole cell lysate at 20 µg

Lane 2:

LRP6 knockout HAP1 whole cell lysate at 20 µg

Lane 3:

HeLa (human cervix adenocarcinoma epithelial cell), whole cell lysate at 20 µg

Lane 4:

HepG2 (human hepatocellular carcinoma epithelial cell), whole cell lysate at 20 µg

Lane 5:

293T (human embryonic kidney epithelial cell), whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Predicted band size: 180 kDa

Observed band size: 180-210 kDa

false

• WB

Supplier Data

Western blot - Anti-LRP6 antibody [EPR22910-39] (AB231779)

The 36 KD band observed is consistent with the dimer of human LRP6 recombinant protein (aa 1476-1600).

All lanes:

Western blot - Anti-LRP6 antibody [EPR22910-39] (ab231779) at 1/1000 dilution

Lane 1:

His-tagged human LRP5 recombinant protein, 10 ng

Lane 2:

His-tagged human LRP6 recombinant protein, 10 ng

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Predicted band size: 180 kDa

Observed band size: 18 kDa,36 kDa

false

Exposure time: 1s

• WB

Lab

Western blot - Anti-LRP6 antibody [EPR22910-39] (AB231779)

False colour image of Western blot : Anti-LRP6 antibody [EPR22910-39] staining at 1/1000 dilution, shown in green; Mouse anti-CANX [CANX/1543] (ab238078) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab231779 was shown to bind specifically to LRP6. A band was observed at 180/200 kDa in wild-type HepG2 cell lysates with no signal observed at this size in LRP6 knockout cell line ab277909 (knockout cell lysate ab284222). To generate this image, wild-type and LRP6 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 5 % 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 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.

All lanes:

Western blot - Anti-LRP6 antibody [EPR22910-39] (ab231779) at 1/1000 dilution

Lane 1:

Wild-type HepG2 cell lysate at 20 µg

Lane 2:

LRP6 knockout HepG2 cell lysate at 20 µg

Lane 2:

Western blot - Human LRP6 knockout Hep G2 cell line (<a href='/en-us/products/cell-lines/human-lrp6-knockout-hep-g2-cell-line-ab277909'>ab277909</a>)

Lane 3:

HeLa cell lysate at 20 µg

Predicted band size: 180 kDa

Observed band size: 180 kDa,200 kDa

false