Anti-LATS1/WARTS antibody [EPR23057-116]

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Anti-LATS1/WARTS antibody [EPR23057-116] (AB243656)

Intracellular flow cytometric analysis of 4% paraformaldehyde fixed, 90% methanol permeabilized HeLa (human cervix adenocarcinoma epithelial cell) cells labelling LATS1/WARTS with ab243656 at 1/500 dilution (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat anti rabbit IgG (Alexa Fluor^® 488, ab150077) at 1/2000 dilution was used as the secondary antibody.

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Anti-LATS1/WARTS antibody [EPR23057-116] (AB243656)

Intracellular flow cytometric analysis of 4% paraformaldehyde, fixed 90% methanol permeabilized NIH/3T3 (mouse embryonic fibroblast) cells labelling LATS1/WARTS with ab243656 at 1/500 dilution (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat anti rabbit IgG (Alexa Fluor^® 488, ab150077) at 1/2000 dilution was used as the secondary antibody.

• WB

Lab

Western blot - Anti-LATS1/WARTS antibody [EPR23057-116] (AB243656)

Blocking and dilution buffer : 5% NFDM/TBST.

Exposure times.

Lane 1 : 81 seconds;

Lanes 2-3 : 3 minutes

The molecular weight observed is consistent with what has been described in the literature (PMID : 30293781).

Bands below 150KD are likely to be degraded LATS1 (PMID : 23573293).

Lysates should be made fresh and used in WB immediately to minimize protein degradation.

All lanes:

Western blot - Anti-LATS1/WARTS antibody [EPR23057-116] (ab243656) at 1/1000 dilution

Lane 1:

NIH/3T3 (mouse embryonic fibroblast) whole cell lysate at 10 µg

Lane 2:

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

Lane 3:

PC-12 (rat adrenal gland pheochromocytoma) whole cell lysate at 10 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 1/100000 dilution

Predicted band size: 126 kDa

Observed band size: 150 kDa

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• WB

Lab

Western blot - Anti-LATS1/WARTS antibody [EPR23057-116] (AB243656)

Western blot : Anti-LATS1 antibody [EPR23057-116] (ab243656) staining at 1/2000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab243656 was shown to bind specifically to LATS1. A band was observed at 127 kDa in wild-type THP-1 cell lysates with no signal observed at this size in LATS1 knockout cell line. To generate this image, wild-type and LATS1 knockout THP-1 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 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.

All lanes:

Western blot - Anti-LATS1/WARTS antibody [EPR23057-116] (ab243656) at 1/2000 dilution

Lane 1:

Wild-type THP-1 cell lysate at 20 µg

Lane 2:

Western blot - Human LATS1 (WARTS) knockout THP-1 cell line (<a href='/en-us/products/cell-lines/human-lats1-warts-knockout-thp-1-cell-line-ab277862'>ab277862</a>)

Lane 2:

LATS1 knockout THP-1 cell lysate at 20 µg

Lane 3:

HT-29 cell lysate at 20 µg

Lane 4:

PC-3 cell lysate at 20 µg

Lane 5:

HeLa cell lysate at 20 µg

Secondary

All lanes:

Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution

Predicted band size: 126 kDa

Observed band size: 127 kDa

false

• WB

Unknown

Western blot - Anti-LATS1/WARTS antibody [EPR23057-116] (AB243656)

Blocking and dilution buffer : 5% NFDM/TBST.

Exposure time : 3 minutes.

Samples loaded are extracts of E.coil expressing either LATS1 or LATS2.

All lanes:

Western blot - Anti-LATS1/WARTS antibody [EPR23057-116] (ab243656) at 1/1000 dilution

Lane 1:

His-tagged human LATS1 recombinant protein at 10 ng

Lane 2:

His-tagged human LATS2 recombinant protein at 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 1/50000 dilution

Predicted band size: 126 kDa

false

Exposure time: 3min