Anti-ATM antibody [EPR17059]

• IP

Supplier Data

Immunoprecipitation - Anti-ATM antibody [EPR17059] (AB199726)

ATM was immunoprecipitated from 1mg of HEK-293 (Human epithelial cells from embryonic kidney) whole cell lysate with ab199726 at 1/100 dilution. Western blot was performed from the immunoprecipitate using ab199726 at 1/1000 dilution. Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG, was used as secondary antibody at 1/1500 dilution.

Lane 1 : HEK-293 whole cell lysate 10ug (Input). Lane 2 : ab199726 IP in HEK-293 whole cell lysate. Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab199726 in HEK-293 whole cell lysate.

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

Exposure time : 10 seconds.

All lanes:

Immunoprecipitation - Anti-ATM antibody [EPR17059] (ab199726)

Predicted band size: 351 kDa

false

• IP

Supplier Data

Immunoprecipitation - Anti-ATM antibody [EPR17059] (AB199726)

ATM was immunoprecipitated from 1mg of SH-SY5Y (Human neuroblastoma from bone marrow cells) whole cell lysate with ab199726 at 1/100 dilution. Western blot was performed from the immunoprecipitate using ab199726 at 1/500 dilution (Panel A) or ab32420 at 1/500 dilution (Panel B). Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG, was used as secondary antibody at 1/1500 dilution.

Lane 1 : SH-SY5Y whole cell lysate 10ug (Input). Lane 2 : ab199726 IP in SH-SY5Y whole cell lysate. Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab199726 in SH-SY5Y whole cell lysate.

Blocking and dilution buffer and concentration : 5% NFDM/TBST.
Exposure time : 30 seconds (Panel A and B).

All lanes:

Immunoprecipitation - Anti-ATM antibody [EPR17059] (ab199726)

Predicted band size: 351 kDa

false

• IP

Supplier Data

Immunoprecipitation - Anti-ATM antibody [EPR17059] (AB199726)

ATM was immunoprecipitated from 1mg of HEK-293 (Human epithelial cells from embryonic kidney) whole cell lysate with ab199726 at 1/100 dilution. Western blot was performed from the immunoprecipitate using ab199726 at 1/500 dilution (Panel A) or ab32420 at 1/500 dilution (Panel B). Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG, was used as secondary antibody at 1/1500 dilution.

Lane 1 : HEK-293 whole cell lysate 10ug (Input). Lane 2 : ab199726 IP in HEK-293 whole cell lysate. Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab199726 in HEK-293 whole cell lysate.

Blocking and dilution buffer and concentration : 5% NFDM/TBST.
Exposure time : 30 seconds (Panel A and B).

All lanes:

Immunoprecipitation - Anti-ATM antibody [EPR17059] (ab199726)

Predicted band size: 351 kDa

false

• WB

Supplier Data

Western blot - Anti-ATM antibody [EPR17059] (AB199726)

Blocking/Dilution buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-ATM antibody [EPR17059] (ab199726) at 1/2000 dilution

Lane 1:

Rat testis lysate at 10 µg

Lane 2:

Mouse testis lysate at 10 µg

Lane 3:

293 (Human epithelial cells from embryonic kidney) whole cell lysate at 10 µg

Lane 4:

SH-SY5Y (Human neuroblastoma from bone marrow cells) whole cell lysate at 10 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution

Predicted band size: 351 kDa

Observed band size: 350 kDa

false

Exposure time: 20s

• WB

Lab

Western blot - Anti-ATM antibody [EPR17059] (AB199726)

False colour image of Western blot : Anti-ATM antibody [EPR17059] staining at 1/2000 dilution, shown in green; Mouse anti-Alpha Tubulin [DM1A] (ab7291) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab199726 was shown to bind specifically to ATM. A band was observed at 350 kDa in wild-type A549 cell lysates with no signal observed at this size in ATM knockout cell line ab276095 (knockout cell lysate ab283834). To generate this image, wild-type and ATM 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 5% 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 (IRDye^® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preabsorbed (ab216776) at 1/20000 dilution.

All lanes:

Western blot - Anti-ATM antibody [EPR17059] (ab199726) at 1/2000 dilution

Lane 1:

Wild-type A549 cell lysate at 20 µg

Lane 2:

ATM knockout A549 cell lysate at 20 µg

Lane 2:

Western blot - Human ATM knockout A549 cell line (<a href='/en-us/products/cell-lines/human-atm-knockout-a549-cell-line-ab276095'>ab276095</a>)

Lane 3:

HEK-293 cell lysate at 20 µg

Lane 4:

U-2 OS cell lysate at 20 µg

Predicted band size: 351 kDa

Observed band size: 350 kDa

false

• WB

Supplier Data

Western blot - Anti-ATM antibody [EPR17059] (AB199726)

Blocking/Dilution buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-ATM antibody [EPR17059] (ab199726) at 1/5000 dilution

All lanes:

293 (Human epithelial cells from embryonic kidney) whole cell lysate at 10 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution

Predicted band size: 351 kDa

Observed band size: 350 kDa

false

Exposure time: 3min

• WB

Supplier Data

Western blot - Anti-ATM antibody [EPR17059] (AB199726)

Blocking/Dilution buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-ATM antibody [EPR17059] (ab199726) at 1/2000 dilution

Lane 1:

Rat testis lysate at 10 µg

Lane 2:

Mouse testis lysate at 10 µg

Lane 3:

Mouse spleen lysate at 10 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution

Predicted band size: 351 kDa

Observed band size: 350 kDa

false

Exposure time: 3min

• WB

CiteAb

Western blot - Anti-ATM antibody [EPR17059] (AB199726)

ATM western blot using anti-ATM antibody [EPR17059] ab199726. Publication image and figure legend from Zhao, X., Guo, X., et al., 2018, Cell Death Dis, PubMed 29352124.

ab199726 was used in this publication in western blot. This may not be the same as the application(s) guaranteed by Abcam. For a full list of applications guaranteed by Abcam for ab199726 please see the product overview.

HBX specifically inhibits CHK2 phosphorylationa, b Western blot for γH2AX in control and HBX-expressing SUDHL-4 a, and DB b, cells with or without MTX treatment (4 ng/ml) for 48 h. c, d Western blot for DDR proteins (P-CHK2/CHK2/ATM and P-CHK1/CHK1) in control and HBX-expressing SUDHL-4 c, and DB d, cells with or without MTX treatment (4 ng/ml) for 48 h. e, f The mRNA levels of DDR proteins (ATM, CHK2, and CHK1) were largely unchanged in control and HBX-expressing SUDHL-4 e, and DB f, cells with or without MTX treatment. g,h The expression of the P-CHK2 downstream genes P53 and P21 in control and HBX-expressing SUDHL-4 g, and DB h, cells with or without MTX treatment. The results are shown as the mean ± SEM from triplicate experiments. GAPDH was used as a loading control

false