Anti-Vimentin (phospho S73) antibody [EP1070Y]

• IP

Unknown

Immunoprecipitation - Anti-Vimentin (phospho S73) antibody [EP1070Y] (AB52944)

ab52944 (purified) at 1 : 20 dilution (2μg) immunoprecipitating Vimentin in HeLa starved 24 hours, then treated with 100nM Calyculin A for 30 minutes whole cell lysate.
Lane 1 (input) : HeLa (Human cervix adenocarcinoma epithelial cell) starved 24 hours,then treated with 100nM Calyculin A for 30 minutes whole cell lysate 10ug
Lane 2 (+) : ab52944 & HeLa starved 24 hours, then treated with 100nM Calyculin A for 30 minutes whole cell lysate
Lane 3 (-) : Rabbit monoclonal IgG (ab172730) instead of ab52944 in HeLa starved 24 hours,then treated with 100nM Calyculin A for 30 minutes whole cell lysate
For western blotting, VeriBlot for IP Detection Reageant (HRP) (ab131366) was used for detection at 1 : 1000 dilution.
Blocking and diluting buffer : 5% NFDM/TBST.

All lanes:

Immunoprecipitation - Anti-Vimentin (phospho S73) antibody [EP1070Y] (ab52944)

Predicted band size: 53 kDa

false

• WB

Supplier Data

Western blot - Anti-Vimentin (phospho S73) antibody [EP1070Y] (AB52944)

Blocking and diluting buffer : 5% NFDM/TBST

All lanes:

Western blot - Anti-Vimentin (phospho S73) antibody [EP1070Y] (ab52944) at 1/10000 dilution

Lane 1:

C6 (Rat glial tumor cell line) whole cell lysate, untreated at 15 µg

Lane 2:

C6 (Rat glial tumor glial cell) starved for 24 hours, then treated with 100ng Calyculin A for 60 minutes, whole cell lysate at 15 µg

Secondary

All lanes:

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

false

• WB

Supplier Data

Western blot - Anti-Vimentin (phospho S73) antibody [EP1070Y] (AB52944)

Blocking and diluting buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-Vimentin (phospho S73) antibody [EP1070Y] (ab52944) at 1/10000 dilution

Lane 1:

Untreated with NIH/3T3 (Mouse embryonic fibroblast) whole cell lysates at 15 µg

Lane 2:

NIH/3T3 (Mouse embryonic fibroblast) treated with 100ng/ml Calyculin A for 30 minutes whole cell lysates at 15 µg

Secondary

All lanes:

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

Predicted band size: 53 kDa

false

• WB

Supplier Data

Western blot - Anti-Vimentin (phospho S73) antibody [EP1070Y] (AB52944)

Blocking and diluting buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-Vimentin (phospho S73) antibody [EP1070Y] (ab52944) at 1/10000 dilution

Lane 1:

Untreated with HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysates at 15 µg

Lane 2:

HeLa (Human cervix adenocarcinoma epithelial cell) starved for 24 hours,then treated with 100nM Calyculin A for 30 minutes whole cell lysates at 15 µg

Secondary

All lanes:

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

Predicted band size: 53 kDa

false

• Dot

Unknown

Dot Blot - Anti-Vimentin (phospho S73) antibody [EP1070Y] (AB52944)

Primary antibody dilution : 1/1000
Secondary antibody : goat anti-rabbit IgG, (H+L), peroxidase conjugated (1/2500 dilution)
Blocking & dilutiing buffer : 5% NFDM/TBST

Lane 1 sample : Vimentin (pS72) phospho peptide
Lane 2 sample : Vimentin non-phospho peptide

Exposure time : 3 minutes