Anti-ATM (phospho S1987) antibody [EPR28058-71]

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-ATM (phospho S1987) antibody [EPR28058-71] (AB315019)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized MEF (mouse embryo fibroblast) cells labelling ATM (phospho S1987) with ab315019 at 1/100 (5.11 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 2 ug/ml dilution (Green). Confocal image showing increased nuclear staining in MEF cells treated with UV-C 100 J/m2, then recovery for 4 hours. 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 2.5ug/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) preadsorbed at 1/1000 2 ug/ml dilution.

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-ATM (phospho S1987) antibody [EPR28058-71] (AB315019)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized MEF (mouse embryo fibroblast) treated with 100 J/m2 UV-C then recovery for 4h (Red) / Untreated MEF (Green) cells labelling ATM (phospho S1987) with ab315019 at 1/500 dilution (0.1 ug)/Red and Green 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, ab150081) at 1/5000 dilution was used as the secondary antibody.

• WB

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Western blot - Anti-ATM (phospho S1987) antibody [EPR28058-71] (AB315019)

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

The identity of the lower MW band at approximately 190 kDa is unknown.

In Western blot, Anti-Vinculin antibody [EPR8185] (ab129002) staining at 1/10000 dilution.

In Western blot, Anti-ATM antibody [EPR20100] - (ab201022) staining at 1/1000 dilution.

All lanes:

Western blot - Anti-ATM (phospho S1987) antibody [EPR28058-71] (ab315019) at 1/1000 dilution

Lane 1:

Untreated NIH/3T3 (mouse embryonic fibroblast) whole cell lysate (untreated membrane) at 20 µg

Lane 2:

NIH/3T3 treated with UV-C for 100J/cm2, then recovery 2 hours, whole cell lysate (untreated membrane) at 20 µg

Lane 3:

NIH/3T3 treated with UV-C for 100J/cm2, then recovery 2 hours, whole cell lysate (phosphatase treated membrane) 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

Observed band size: 349 kDa,124 kDa

false

Exposure time: 81s

• WB

Supplier Data

Western blot - Anti-ATM (phospho S1987) antibody [EPR28058-71] (AB315019)

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

The identity of the lower MW band at approximately 190 kDa is unknown.

In Western blot, Anti-Vinculin antibody [EPR8185] (ab129002) staining at 1/10000 dilution.

In Western blot, Anti-ATM antibody [EPR20100] - (ab201022) staining at 1/1000 dilution.

All lanes:

Western blot - Anti-ATM (phospho S1987) antibody [EPR28058-71] (ab315019) at 1/1000 dilution

Lane 1:

Untreated MEF (mouse embryo fibroblast) whole cell lysate (untreated membrane) at 20 µg

Lane 2:

MEF treated with UV-C for 100J/m2, then recovery 4 hours, whole cell lysate (untreated membrane) at 20 µg

Lane 3:

MEF treated with UV-C for 100J/m2, then recovery 4 hours, whole cell lysate (phosphatase treated membrane) 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

Observed band size: 349 kDa,124 kDa

false

Exposure time: 81s

• WB

Supplier Data

Western blot - Anti-ATM (phospho S1987) antibody [EPR28058-71] (AB315019)

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

The identity of the lower MW band at approximately 190 kDa and higher MW band at approximately 460 kDa are unknown.

In Western blot, Anti-Vinculin antibody [EPR8185] (ab129002) staining at 1/10000 dilution.

In Western blot, Anti-6X His tag® antibody [EPR20547] - ChIP Grade (ab213204) staining at 1/5000 dilution.

All lanes:

Western blot - Anti-ATM (phospho S1987) antibody [EPR28058-71] (ab315019) at 1/1000 dilution

Lane 1:

293T cells transfected with an empty vector containing a myc-His-tag®, whole cell lysate at 20 µg

Lane 2:

293T cells transfected with a mouse wild-type ATM expression vector containing a myc-His-tag®, whole cell lysate at 20 µg

Lane 3:

293T cells transfected with a mouse ATM (S1987A mutation) expression vector containing a myc-His-tag®, 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

Observed band size: 349 kDa,124 kDa

false

Exposure time: 180s

• Dot

Supplier Data

Dot Blot - Anti-ATM (phospho S1987) antibody [EPR28058-71] (AB315019)

Dot blot analysis of ATM (phospho S1987) using ab315019 at 1 : 1000 (0.511 ug/ml) followed by a Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1 : 100,000 dilution.

Exposure time : 180 seconds

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