Anti-ATM antibody [Y170]

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ATM antibody [Y170] (AB32420)

Formaldehyde-fixed human colon tissue stained for ATM using ab32420 at 1/100 dilution in immunohistochemical analysis.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ATM antibody [Y170] (AB32420)

Immunohistochemical analysis of formalin fixed paraffin embedded human breast carcinoma labelling ATM with ab32420 at a concentration of 0.1µg/ml. The immunostaining was performed on a Ventana DISCOVERY ULTRA (Roche Tissue Diagnostics) instrument with a OptiView DAB IHC Detection Kit. Heat mediated antigen retrieval was performed with DISCOVERY cell conditioning solution (CC1) 100°C, pH8.5 for 32mins. ab32420 anti-ATM antibody [Y170] was incubated for 16mins at 37°C. Sections were counterstained with Hematoxylin II. Image inset shows absence of staining in secondary antibody only control.

Customers are encouraged to optimise antigen retrieval conditions, antibody concentration, incubation times and temperature for best results in their own IHC assay workflow (automated and manual).

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ATM antibody [Y170] (AB32420)

Immunohistochemical analysis of paraffin-embedded Human breast cancer tissue labeling ATM with ab32420 at 1 : 100 dilution. Tissue underwent antigen retrieval using Tris/EDTA Buffer (pH9.0). The section was counterstained with haematoxylin.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ATM antibody [Y170] (AB32420)

Immunohistochemical analysis of paraffin-embedded Human normal breast tissue labeling ATM with ab32420 at 1 : 100 dilution. Tissue underwent antigen retrieval using Tris/EDTA Buffer (pH9.0). The section was counterstained with haematoxylin.

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-ATM antibody [Y170] (AB32420)

Immunocytochemistry/ Immunofluorescence analysis of HeLa cells labeling ATM with ab32420 at 1/500. Goat anti rabbit IgG(Alexa Fluor^® 488), ab150077 at 1/1000 was used as the secondary antibody. Cells were fixed with 4% Paraformaldehyde and permeabilised with 0.1% tritonX-100. Nulei were counterstained blue with DAPI.

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-ATM antibody [Y170] (AB32420)

Immunocytochemistry/ Immunofluorescence analysis of HepG2 cells labeling ATM with ab32420 at 1/500. Goat anti rabbit IgG(Alexa Fluor^® 488), ab150077 at 1/1000 was used as the secondary antibody. Cells were fixed with 4% Paraformaldehyde and permeabilised with 0.1% tritonX-100. Nulei were counterstained blue with DAPI.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ATM antibody [Y170] (AB32420)

Formaldehyde-fixed human serous ovarian tumor tissue stained for ATM using ab32420 at 1/50 dilution in immunohistochemical analysis.

• Flow Cyt (Intra)

Unknown

Flow Cytometry (Intracellular) - Anti-ATM antibody [Y170] (AB32420)

Intracellular Flow Cytometry analysis of HeLa (human cervix adenocarcinoma) cells labelling ATM with purified ab32420 at 1/20 (red). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. An Alexa Fluorr^®488-conjugated goat anti-rabbit IgG (1/2000) was used as the secondary antibody. Black - Isotype control, rabbit monoclonal IgG. Blue - Unlabelled control, cells without incubation with primary and secondary antibodies.

• WB

Lab

Western blot - Anti-ATM antibody [Y170] (AB32420)

All lanes:

Western blot - Anti-ATM antibody [Y170] (ab32420) at 1/3000 dilution

Lane 1:

HeLa cell lysate at 10 µg

Lane 2:

HEK293 cell lysate at 10 µg

Secondary

All lanes:

HRP goat anti-rabbit (H+L) at 1/1000 dilution

Predicted band size: 351 kDa

Observed band size: 370 kDa

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• ICC/IF

PubMed

Immunocytochemistry/ Immunofluorescence - Anti-ATM antibody [Y170] (AB32420)

DNA repair proteins accumulate at MVM APAR bodies

Repair proteins accumulate at APAR bodies. NB324K cells were infected with MVMp (MOI of 10) for 16 hr before being fixed and processed for immunofluorescence. Cells were stained with the indicated antibodies to mark DDR repair proteins. APAR bodies were detected with antibodies to NS1. Nuclei were stained with DAPI. All images were captured using an objective of 63×.

Cells were fixed with 4% paraformaldehyde for 15 minutes and permeabilized with 0.5% Triton X-100 in PBS for 15 minutes.

(Image shows the right-hand panel of Figure 2A)

Image from Adeyemi R.O. et al PLoS Pathog. 2010 Oct 7;6(10):e1001141. doi: 10.1371/journal.ppat.1001141.

• WB

Lab

Western blot - Anti-ATM antibody [Y170] (AB32420)

False colour image of Western blot : Anti-ATM antibody [Y170] staining at 1/1000 dilution, shown in green; Mouse anti-Alpha Tubulin [DM1A] (ab7291) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab32420 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 [Y170] (ab32420) at 1/1000 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 [Y170] (AB32420)

Western blot : Anti-ATM antibody [Y170] (ab32420) staining at 1/1000 dilution, shown in green; Mouse anti-CANX [CANX/1543] (ab238078) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab32420 was shown to bind specifically to ATM. A band was observed at 350 kDa in wild-type MCF7 cell lysates with a reduction in signal observed at this size in ATM heterozygous knockout cell line ab282630. To generate this image, wild-type and ATM heterozygous knockout MCF7 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-ATM antibody [Y170] (ab32420) at 1/1000 dilution

Lane 1:

Wild-type MCF7 cell lysate at 20 µg

Lane 2:

ATM knockout MCF7 cell lysate at 20 µg

Lane 3:

Wild-type A549 cell lysate at 20 µg

Lane 4:

ATM knockout A549 ab283811 cell lysate at 20 µg

Observed band size: 350 kDa

false