Western Blot - Anti-ATM (phospho Ser1981) Antibody (A10862)
Western blot analysis of extracts of various cell lines, using Anti-ATM (phospho Ser1981) Antibody (A10862) at 1:1,000 dilution. 293T cells were treated by Doxorubicin (0. 5 uM) at 37°C for 24 hours. 293T cells were treated by UV at room temperature for 15-30 minutes. NIH/3T3 cells were treated by UV at room temperature for 15-30 minutes. The secondary antibody was Goat Anti-Rabbit IgG H&L Antibody (HRP) at 1:10,000 dilution. Lysates/proteins were present at 25µg per lane. The blocking buffer used was 3% non-fat dry milk in TBST. Detection was with a ECL Basic Kit. Exposure time: 180s.
Isabelle Cristine da Costa, University of Manchester
Verified Customer
Applications: Western Blot
Species: Human
Sample: U2OS osteosarcoma cell line
Experimental Conditions: WB: 3-8% TA gel, PVDF, wet transfer 150 mA overnight 4°C; 50 µg loading; lysis RIPA; blocking 1% BSA/TBST 30 min RT; primary 1:500 O/N 4°C; secondary Invitrogen G-21234 1:5000 1 h RT; detection ChemiDoc ECL.
Dilution: WB: 1:500
Results Summary: Detects pATM (S1981) when DNA damage present (IR-treated cells); Signal absent with ATM inhibitor (ATMi); Specific to pATM in irradiated cells.
Comments: Strong, specific signal; clear distinction between IR-treated and ATMi-treated samples.
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