Anti-ATM antibody [2C1 (1A1)] - BSA and Azide free

• WB

Supplier Data

Western blot - Anti-ATM antibody [2C1 (1A1)] - BSA and Azide free (AB78)

5% SDS-PAGE.

Running conditions : 80V, 15min; 140V, 40 minutes.

Transfer condition : Semi-dry, 18 V, 60 min (NC membrane).

Blocking condition : 5% non-fat milk in TBST, RT, 60 minutes.

Primary antibody incubation : 4°C, overnight.

Washing condition : 5 ml TBST, 4 x 5 minutes.

Exposure : enhanced ECL

All lanes:

Western blot - Anti-ATM antibody [2C1 (1A1)] - BSA and Azide free (ab78) at 1/500 dilution

Lane 1:

30ug HeLa

Lane 2:

30ug HeLa nuclear extract

Lane 3:

30ug SK-N-SH

Secondary

All lanes:

Mouse IgG antibody (HRP) at 1/5000 dilution

Predicted band size: 351 kDa

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• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ATM antibody [2C1 (1A1)] - BSA and Azide free (AB78)

ab78 staining ATM in Human Kidney sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Samples were incubated with primary antibody (5ug/ml) and a Biotin-conjugated rabbit anti-mouse IgG was used as the secondary antibody.

• Flow Cyt

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Flow Cytometry - Anti-ATM antibody [2C1 (1A1)] - BSA and Azide free (AB78)

Overlay histogram showing HeLa cells stained with ab78 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab78, 1μg/1x10⁶ cells) for 30 min at 22°C. The secondary antibody used was DyLight^® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2μg/1x10⁶ cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in HeLa cells fixed with 4% paraformaldehyde (10 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ATM antibody [2C1 (1A1)] - BSA and Azide free (AB78)

ab78 (2μg/ml) staining ATM in human colonic mucosa, using an automated system (DAKO Autostainer Plus). Using this protocol there is strong nuclear staining of mucosal epithelium and lymphocytes.
Sections were rehydrated and antigen retrieved with the Dako 3 in 1 AR buffer EDTA pH 9.0 in a DAKO PT link. Slides were peroxidase blocked in 3% H2O2 in methanol for 10 mins. They were then blocked with Dako Protein block for 10 minutes (containing casein 0.25% in PBS) then incubated with primary antibody for 20 min and detected with Dako envision flex amplification kit for 30 minutes. Colorimetric detection was completed with Diaminobenzidine for 5 minutes. Slides were counterstained with Haematoxylin and coverslipped under DePeX. Please note that, for manual staining, optimization of primary antibody concentration and incubation time is recommended. Signal amplification may be required.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ATM antibody [2C1 (1A1)] - BSA and Azide free (AB78)

ab78 staining ATM in Human Testis sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Samples were incubated with primary antibody (5ug/ml) and a Biotin-conjugated rabbit anti-mouse IgG was used as the secondary antibody.

• IP

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Immunoprecipitation - Anti-ATM antibody [2C1 (1A1)] - BSA and Azide free (AB78)

Detection of human ATM protein using anti-ATM 2C1 monoclonal antibody (ab78) by immunoprecipitation.

All lanes:

Immunoprecipitation - Anti-ATM antibody [2C1 (1A1)] - BSA and Azide free (ab78)

Predicted band size: 351 kDa

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• WB

Supplier Data

Western blot - Anti-ATM antibody [2C1 (1A1)] - BSA and Azide free (AB78)

Detection of human ATM protein using anti-ATM 2C1 monoclonal antibody (ab78) by western blot.

All lanes:

Western blot - Anti-ATM antibody [2C1 (1A1)] - BSA and Azide free (ab78)

Predicted band size: 351 kDa

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• WB

CiteAb

Western blot - Anti-ATM antibody [2C1 (1A1)] - BSA and Azide free (AB78)

ATM western blot using anti-ATM antibody [2C1 (1A1)] - BSA and Azide free ab78. Publication image and figure legend from Cilli, D., Mirasole, C., et al., 2014, PLoS One, PubMed 25485873.

ab78 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 ab78 please see the product overview.

Evaluation of the Strep-tag recombinant proteins expression and Western blot analysis of NBN, p26 and p70 interactors.(A) Protein lysates, obtained after 24 and 48 h from HEK293 transient tranfections, were analyzed by Western blot using a StrepMAB-Classic horse radish peroxidase conjugated antibody. The highest level of expression was observed after 48 h from transfection, for all the recombinant proteins. Protein extracts obtained from HEK293 cells transfected with the empty vector were used as negative control. (B) Check of the expression of the Strep-tag recombinant proteins. Immunoblots were performed using the protein eluates obtained from HEK293 transfected cells either untreated or exposed to 2 Gy of X-rays, lysed after 30 minutes and purified by Strep-tag chromatography. Filters were probed with the anti-NBN antibody directed against the full-length protein. (C) Protein eluates obtained from HEK293 transfected cells exposed to 2 Gy of X-rays were lysed after 30 minutes, purified by Strep-tag chromatography, and analysed by Western blot using the following antibodies : anti-53BP1 rabbit polyclonal, anti-ATM mouse monoclonal, anti-ATM pSer1981 mouse monoclonal, anti-BRCA1 mouse monoclonal, anti-CHK2 pThr68 rabbit polyclonal, anti-CHK2 mouse monoclonal, anti-CtIP rabbit polyclonal, γ-H2AX rabbit polyclonal, anti-Hsp90 rabbit polyclonal, anti-MRE11 mouse monoclonal, anti-NBN mouse monoclonal, anti-PARP1 mouse monoclonal, anti-PP2A rabbit polyclonal, anti-PML rabbit polyclonal, anti-SMC1 rabbit polyclonal.

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