ATM antibody [2C1 (1A1)] (ab78)

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review by Vasileia Sapountzi submitted on 22 June 2004
Application:	Western blot
ATM antibody [2C1 (1A1)] (ab78)
Additional Data
Notes	Ab78 detects very well endogenous ATM from MCF7 whole cell lysates and nuclear extracts in western blots.
Dilution notes	1:500 in TBS-Tween20 Proteins resolved in 6% SDS PAGE and transfered onto nitrocellulose paper (overnight wet transfer (30V) in Tris-glycine buffer without methanol)

review by Stefano Ferrari submitted on 15 April 2002
Application:	Western blot
ATM antibody [2C1 (1A1)] (ab78)
Additional Data
Notes	Detection of ATM with Ab-78 Western blot analysis was carried out on total cell extracts (30 micrograms of total protein) derived from exponentially growing HCT116 (human colon carcinoma) and HeLa (human cervical carcinoma) cells. Ab-78 (Gentex) revealed a single band corresponding to ATM. In the case of MRC5 (human lung fibroblasts) the signal resulted much weaker. The antibody was not capable of immunoprecipitating ATM.
Dilution notes	Cell extraction / immunoprecipitation buffer (buffer A): 50 mM Tris-HCl, pH 7.5; 120 mM NaCl; 20 mM NaF; 1 mM EDTA; 6 mM EGTA; 15 mM Na-pyrophosphate; 1 mM benzamidine; 0.5 mM Na-orthovanadate; 0.1 mM phenylmethylsulfonyl fluoride (PMSF); 1% NP40 Cell extraction was performed using a ratio of 50-100 microliter buffer A / 1 x 10exp6 cells Cell extracts were resolved on 6% SDS-PAGE and proteins transfered onto PVDF using a semi-dry transfer system. Membranes were stained with 5% Ponceau red, destained and blocked with 5% milk in TBS containing 01% Tween-20. Western blot analysis was performed using the following antibody dilution: primary 1/1000; secondary (HRP-linked) 1/5000. Protein bands were revealed with the ECL system. .

review by Randi Syljuåsen submitted on 11 July 2001
Application:	Western blot
ATM antibody [2C1 (1A1)] (ab78)
Additional Data
Notes	The Ab seemed to work fine for Western blotting, however, we got some non-specific activities when we tried to use this antibody for immunoprecipitation followed by in vitro kinase assays. We are therefore looking for better Ab to ATM to use for kinase assays.

review by Anouk Waeytens submitted on 24 April 2001
Application:	Primary Cell Culture
ATM antibody [2C1 (1A1)] (ab78)
Additional Data
Notes	very nice signal in Western blot and immunofluorescence; very pure antibody - no cross-reactivity seen yet

review by Anouk Waeytens submitted on 28 March 2001
Application:	Western blot
ATM antibody [2C1 (1A1)] (ab78)
Additional Data
Notes	I used the ATM - 2C1 antibody in confocal immunocytochemistry (secondary antibody: FITC anti-mouse); but without results. I will continue these experiments. In Western blotting of HeLa, SK-BR-3 and HCC cellysates, the antibody gave nice results and it appears to be rather specific. Further Western blotting will be carried out with this antibody.