Anti ATM Protein Kinase, phospho Ser1981

Anti ATM Antibody showing overlay of anti ATM pS1981 staining. Cells were fixed 15 min after 5 Gy (IR+) of irradiation, then labeled with antibody. See Kitagawa et al. for additional details.

Anti ATM Antibody showing overlay of anti-ATM pS1981 staining. Cells were fixed 15 min after 5 Gy (IR+) of irradiation, then labeled with antibody. See Kitagawa et al. for additional details.

Rockland's anti-ATM pS1981 mouse monoclonal antibody (Catalog # 200-301-400) detects ATM phosphorylated on Ser 1981 by Indirect immunofluorescence microscopy. Shown are hTCEpi cells (courtesy of Dr. Danielle Robertson) infected with HSV-1 at MOI 5.0 and fixed at 8 hpi with 3% paraformaldehyde/2% sucrose for 10 min. After rinsing, cells were permeabilized with 0.5% Triton X-100 for 5 min, blocked with 3% BSA for 30 min, and stained with Rockland's primary anti-ATM pS1981 antibody overnight at 5 ug/mL (1:200). Secondary staining was performed with Alexa Fluor 594 anti-mouse antibody. Images were taken with Olympus AX70 compound epifluorescence microscope equipped with Spot RT Slider camera. Experiment was performed by Oleg Alekseev in the laboratory of Dr. Jane Azizkhan-Clifford at Drexel University College of Medicine.

Western Blot of Rockland's Protein A Purified Mab anti-ATM Protein Kinase pS1981. Lane 1: HEK293 cells treated with doxorubicin pre-incubation of peptide with 50 ug of immunizing phospho peptide negates specific staining. Lane 2: HEK293 cells treated with doxorubicin. A 370 kDa band corresponding to phosphorylated ATM is detected (lane 2). The lysate was prepared with HALT phosphatase inhibitor (Pierce). Load: ~30ug. Primary antibody: anti-ATM Protein Kinase pS1981 diluted 1:500 overnight at 4C. Secondary Antibody: IRDyeTM800 conjugated Gt-a-Mouse IgG [H&L] (code 610-132-121) at 1:10,000 for 40 min at room temperature. LICOR's OdysseyR Infrared Imaging System was used to scan and process the image. Other detection systems will yield similar results.