重组Anti-LATS2抗体[EPR23126-488] - BSA and Azide free

• IP

Supplier Data

Immunoprecipitation - Anti-LATS2 antibody [EPR23126-488] - BSA and Azide free (AB280363)

This data was developed using ab243657, the same antibody clone in a different buffer formulation.

LATS2 was immunoprecipitated from 0.35 mg HeLa (human cervix adenocarcinoma epithelial cell), whole cell lysate with ab243657 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab243657 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP)(ab131366) was used at 1/5000 dilution.

Lane 1 : HeLa (human cervix adenocarcinoma epithelial cell), whole cell lysate 10 ug

Lane 2 : ab243657 IP in HeLa whole cell lysate

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab243657 in HeLa whole cell lysate

Blocking and dilution buffer and concentration : 5% NFDM/TBST.

Exposure time : 64 seconds

Fresh lysates were used in this WB.

All lanes:

Immunoprecipitation - Anti-LATS2 antibody [EPR23126-488] (<a href='/products/primary-antibodies/lats2-antibody-epr23126-488-ab243657'>ab243657</a>)

Predicted band size: 120 kDa

Observed band size: 140 kDa

false

• WB

Lab

Western blot - Anti-LATS2 antibody [EPR23126-488] - BSA and Azide free (AB280363)

This data was developed using ab243657, the same antibody clone in a different buffer formulation.

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

Fresh lysates were used in this WB.

This blot was developed using a higher sensitivity ECL substrate.

Exposure time : 3 minutes

All lanes:

Western blot - Anti-LATS2 antibody [EPR23126-488] (<a href='/products/primary-antibodies/lats2-antibody-epr23126-488-ab243657'>ab243657</a>) at 1/1000 dilution

Lane 1:

HeLa (human cervix adenocarcinoma epithelial cell), transfected with scrambled siRNA control, whole cell lysate at 20 µg

Lane 2:

HeLa transfected with siRNA specifically targeting LATS2, whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Predicted band size: 120 kDa

Observed band size: 140 kDa

false

• WB

Lab

Western blot - Anti-LATS2 antibody [EPR23126-488] - BSA and Azide free (AB280363)

This data was developed using ab243657, the same antibody clone in a different buffer formulation.

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

Fresh lysates were used in this WB.

Exposure time : 3 minutes

All lanes:

Western blot - Anti-LATS2 antibody [EPR23126-488] (<a href='/products/primary-antibodies/lats2-antibody-epr23126-488-ab243657'>ab243657</a>) at 1/1000 dilution

Lane 1:

HeLa (human cervix adenocarcinoma epithelial cell), whole cell lysate at 10 µg

Lane 2:

HEK-293T (human embryonic kidney epithelial cell), whole cell lysate at 10 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Predicted band size: 120 kDa

Observed band size: 140 kDa

false

• WB

Lab

Western blot - Anti-LATS2 antibody [EPR23126-488] - BSA and Azide free (AB280363)

This data was developed using ab243657, the same antibody clone in a different buffer formulation.

Western blot : Anti-LATS2 antibody [EPR23126-488] (ab243657) staining at 1/1000 dilution, shown in green; Mouse anti-CANX [CANX/1543] (ab238078) loading control staining at 1/20000 dilution, shown in magenta. In Western blot, ab243657 was shown to bind specifically to LATS2. A band was observed at 117-171 kDa in wild-type A549 cell lysates with no signal observed at this size in LATS2 knockout cell line. To generate this image, wild-type and LATS2 knockout A549 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-LATS2 antibody [EPR23126-488] (<a href='/products/primary-antibodies/lats2-antibody-epr23126-488-ab243657'>ab243657</a>) at 1/1000 dilution

Lane 1:

Wild-type A549 cell lysate at 20 µg

Lane 2:

Western blot - Human LATS2 knockout A549 cell line (<a href='/products/cell-lines/human-lats2-knockout-a549-cell-line-ab286746'>ab286746</a>)

Lane 2:

LATS2 knockout A549 cell lysate at 20 µg

Lane 3:

MCF7 cell lysate at 20 µg

Lane 4:

Raji cell lysate at 20 µg

Secondary

Lanes 1 - 4:

Goat anti-Rabbit IgG H&L 800CW at 1/20000 dilution

Lanes 1 - 4:

Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution

Observed band size: 117-171 kDa

false

• WB

Lab

Western blot - Anti-LATS2 antibody [EPR23126-488] - BSA and Azide free (AB280363)

This data was developed using ab243657, the same antibody clone in a different buffer formulation.

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

Exposure time : 3 minutes

All lanes:

Western blot - Anti-LATS2 antibody [EPR23126-488] (<a href='/products/primary-antibodies/lats2-antibody-epr23126-488-ab243657'>ab243657</a>) at 1/1000 dilution

Lane 1:

His-tagged human LATS1 recombinant protein (aa701-1054) at 0.01 µg

Lane 2:

His-tagged human LATS2 recombinant protein (aa664-1016) at 0.01 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Predicted band size: 120 kDa

false