Anti-LATS1/WARTS (phospho T1079) + LATS2 (phospho T1041) antibody [EPR27261-8]

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-LATS1/WARTS (phospho T1079) + LATS2 (phospho T1041) antibody [EPR27261-8] (AB305029)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized HeLa (human cervical adenocarcinoma epithelial cell) cells labelling LATS1(phospho T1079)+LATS2 (phospho T1041) with ab305029 at 1/100 (5.0 µg/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preabsorbed antibody at 1/1000 2 µg/ml dilution (Green). Confocal image showing mainly centrosome staining in HeLa cells treated with Calyculin A (100 ng/ml) for 10 min. Image was taken with a confocal microscope(Leica-Microsystems, TCS SP8). ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 2.5µg/ml dilution (Red). The Nuclear counterstain was DAPI (Blue). Secondary antibody only control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preabsorbed at 1/1000 2 µg/ml dilution.

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-LATS1/WARTS (phospho T1079) + LATS2 (phospho T1041) antibody [EPR27261-8] (AB305029)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized HeLa (human cervical adenocarcinoma epithelial cell) cells labelling LATS1(phospho T1079)+LATS2 (phospho T1041) with ab305029 at 1/100 (5.0 µg/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preabsorbed antibody at 1/1000 2 µg/ml dilution (Green). Confocal image showing centriole (arrow) staining in HeLa cells in M phase. Image was taken with a confocal microscope(Leica-Microsystems, TCS SP8). ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 2.5µg/ml dilution (Red). The Nuclear counterstain was DAPI (Blue). Secondary antibody only control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preabsorbed at 1/1000 2 µg/ml dilution.

• WB

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Western blot - Anti-LATS1/WARTS (phospho T1079) + LATS2 (phospho T1041) antibody [EPR27261-8] (AB305029)

Blocking and diluting buffer and concentration : 5% NFDM/TBST The antibody detects human LATS1 (phospho T1079) & LATS2 (phospho T1041). In Western blot, anti-His antibody (ab213204) staining at 1/5000 dilution. Exposure time : Lanes 1-2 : 41 seconds, lanes 3-4 and 7-8 : 180 seconds, lanes 5-6 : 10 seconds.

All lanes:

Western blot - Anti-LATS1/WARTS (phospho T1079) + LATS2 (phospho T1041) antibody [EPR27261-8] (ab305029) at 1/1000 dilution

Lanes 1 - 2:

HEK293T cells transfected with a human LATS1(S909A mutation) expression vector containing a his tag

Lanes 3 - 4:

HEK293T cells transfected with a human LATS1(T1079A mutation) expression vector containing a his tag

Lanes 5 - 6:

HEK293T cells transfected with a human LATS2(S872A mutation) expression vector containing a his tag

Lanes 7 - 8:

HEK293T cells transfected with a human LATS2(T1041A mutation) expression vector containing a his tag

Secondary

All lanes:

Western blot - VeriBlot for IP Detection Reagent (HRP) (<a href='/en-us/products/reagents/veriblot-for-ip-detection-reagent-hrp-ab131366'>ab131366</a>) at 1/5000 dilution

Observed band size: 150 kDa

false

• WB

Supplier Data

Western blot - Anti-LATS1/WARTS (phospho T1079) + LATS2 (phospho T1041) antibody [EPR27261-8] (AB305029)

Blocking and diluting buffer and concentration : 5% NFDM/TBST Lysates were freshly made and used immediately to minimize protein degradation. In Western blot, anti-LATS1 antibody (ab243656) staining at 1/1000 dilution. Anti-LATS2 antibody (ab243657) staining at 1/1000 dilution. Anti-GAPDH antibody (ab181602) loading control staining at 1/200000 dilution. Exposure time : 15 seconds

All lanes:

Western blot - Anti-LATS1/WARTS (phospho T1079) + LATS2 (phospho T1041) antibody [EPR27261-8] (ab305029) at 1/1000 dilution

Lane 1:

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

Lane 2:

HeLa treated with 100 ng/mL Calyculin A for 10 minutes (untreated membrane)

Lane 3:

Untreated HeLa whole cell lysate 20 μg (phosphatase treated membrane) at 20 µg

Lane 4:

HeLa treated with 100 ng/mL Calyculin A for 10 minutes (phosphatase treated membrane)

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

Observed band size: 150 kDa

false

Exposure time: 15s

• Dot

Supplier Data

Dot Blot - Anti-LATS1/WARTS (phospho T1079) + LATS2 (phospho T1041) antibody [EPR27261-8] (AB305029)

Dot blot analysis of LATS1(phospho T1079)+LATS2 (phospho T1041 using ab305029 at 1 : 1000 (0.5 ug/ml) followed by a Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1 : 100,000 dilution.

Lane 1 : LATS1 (phospho T1079)/ LATS2 (phospho T1041) peptide a

Lane 2 : LATS1 (phospho T1079)/ LATS2 (phospho T1041) peptide b

Lane 3 : LATS1/ LATS2 non-phospho peptide a

Peptide a, b and c share 100% sequence similarity with human LATS1 and LATS2.

Exposure time : 180 seconds

Blocking and diluting buffer and concentration : 5% NFDM/TBST