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Total atm

Manufactured by Cell Signaling Technology
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Total ATM is a lab equipment product that measures the total levels of the ATM (Ataxia-Telangiectasia Mutated) protein in samples. It provides a quantitative assessment of the ATM protein, which is a key regulator of the cellular response to DNA damage.

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Lab products found in correlation

P21 waf1 cip1, by Cell Signaling Technology (2 mentions) Survivin, by Cell Signaling Technology (2 mentions) Mammalian achaete scute complex like1 ascl1, by BD (2 mentions) Glyceraldehyde 3 phosphate dehydrogenase gapdh, by Bio-Techne (2 mentions) Chromogranin a, by Zymo Research (2 mentions) Chemiluminescence substrate, by Bio-Rad (1 mentions) X omat 2000a, by Kodak (1 mentions) Pchk2 t68, by Cell Signaling Technology (1 mentions) P atm s1981, by Cell Signaling Technology (1 mentions) P p53 s15, by Cell Signaling Technology (1 mentions) Pchk1 s345, by Cell Signaling Technology (1 mentions) Total atr, by Cell Signaling Technology (1 mentions) Patr s428, by Cell Signaling Technology (1 mentions) Total chk2, by Cell Signaling Technology (1 mentions) Vinculin, by Abcam (1 mentions) Cleaved caspase 3, by Cell Signaling Technology (1 mentions) β actin, by Merck Group (1 mentions) Phospho atm, by Cell Signaling Technology (1 mentions) Vimentin, by Merck Group (1 mentions) Rad50, by Abcam (1 mentions)

4 protocols using total atm

1

Immunoblot Analysis of Cell Lysates

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Total BON cell lysates following dacarbazine ± ABT-888 treatment were prepared and analyzed by Western blotting as previously described44 (link). Each antibody was diluted as follows: 1:2000 for mammalian achaete-scute complex-like1 (ASCL1) (BD Pharmingen, San Diego, CA, USA), 1:3000 for chromogranin A (Zymed Laboratories, San Francisco, CA, USA), 1:10,000 for glyceraldehyde-3-phosphate dehydrogenase (GAPDH) (Trevigen, Gaithersburg, MD, USA), and 1:1000 for p21Waf1/Cip1, cleaved poly (ADP-ribose) polymerase (PARP), phosphorylated ATM, total ATM, and Survivin (Cell Signaling Technology, Beverly, MA, USA). Antibody signals were detected using Supersignal West Femto, Dura, or Pico (Pierce, Rockford, IL, USA) chemiluminescence systems and manufacturers’ instructions were adhered to.
Somnay Y., Lubner S., Gill H., Matsumura J.B, & Chen H. (2016). The PARP inhibitor ABT-888 potentiates darbazine-induced cell death in carcinoids. Cancer gene therapy, 23(10), 348-354.
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2

Comprehensive Western Blot Analysis

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Western blot analysis were done with the following antibodies: p-CHK1 S345 (#2348, 1:2,000, Cell Signaling), total CHK1 (#04-207, 1:1,000, Millipore), p-CHK2 T68 (#2197, 1:2,000, Cell Signaling), total CHK2 (#6334, 1:1,000, Cell Signaling), p-ATM S1981 (#5883, 1:1,000, Cell Signaling), total ATM (#2873, 1:1,000, Cell Signaling), p-ATR S428 (#2853, 1:1,000, Cell Signaling), total ATR (#2790, 1:1,000, Cell Signaling), p-p53 S15 (#9286, 1:2,000, Cell Signaling), total p53 (#OP03, AB-1, 1:5,000, Oncogene), cleaved Caspase3 (#9661, 1:2,000, Cell Signaling), PARP (#9542, 1:10,000, Cell Signaling), β-actin (#A5316, 1:10,000, Sigma Aldrich), Vinculin (3AB73412, 1:5,000, Abcam). Protein bands were visualized using chemiluminescence substrates (#170-5061, BioRad) and imaged on Kodak X-OMAT 2000A.
Kim H., Lin Q, & Yun Z. (2018). The hypoxic tumor microenvironment in vivo selects the tumor cells with increased survival against genotoxic stresses. Cancer letters, 431, 142-149.
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3

Immunoblot Analysis of Cell Lysates

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Total BON cell lysates following dacarbazine ± ABT-888 treatment were prepared and analyzed by Western blotting as previously described44 (link). Each antibody was diluted as follows: 1:2000 for mammalian achaete-scute complex-like1 (ASCL1) (BD Pharmingen, San Diego, CA, USA), 1:3000 for chromogranin A (Zymed Laboratories, San Francisco, CA, USA), 1:10,000 for glyceraldehyde-3-phosphate dehydrogenase (GAPDH) (Trevigen, Gaithersburg, MD, USA), and 1:1000 for p21Waf1/Cip1, cleaved poly (ADP-ribose) polymerase (PARP), phosphorylated ATM, total ATM, and Survivin (Cell Signaling Technology, Beverly, MA, USA). Antibody signals were detected using Supersignal West Femto, Dura, or Pico (Pierce, Rockford, IL, USA) chemiluminescence systems and manufacturers’ instructions were adhered to.
Somnay Y., Lubner S., Gill H., Matsumura J.B, & Chen H. (2016). The PARP inhibitor ABT-888 potentiates darbazine-induced cell death in carcinoids. Cancer gene therapy, 23(10), 348-354.
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4

Immunoblotting Assay for Protein Expression

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Cell lysates are prepared using Ripa Buffer and run in SDS-PAGE gel as per standard protocol. After the proteins are transferred in nitrocellulose membrane, blocking was done using 5% BSA. Primary and Secondary antibodies are mixed in 5% BSA. We used primary antibodies of EpCAM (Santa Cruz, sc-25308), phosphoAKT (Sigma Aldrich, SAB5600064), Total AKT (Sigma Aldrich, SAB5600066), Rad50 (Abcam, ab8913). Ku80 (Cell Signaling Technology, CST2180), phosphoATM (Cell Signaling Technology, CST D6H9), Total ATM (Cell Signaling Technology, CST2873), CHK2 (Cell Signaling Technology, CSTC13C1), Vimentin (Sigma Aldrich V6630), Twist (Santa Cruz, sc-15393) and Tubulin (Abcam, ab7291). Primary antibody of EpCAM has dilution of 1:200, while rests of the antibodies have dilution of 1:1000. We used anti-mouse HRP secondary antibody (ab6728) and anti-rabbit secondary antibody (Thermo Scientific 35512) at 1:10000 dilution.
Mal A., Bukhari A.B., Singh R.K., Kapoor A., Barai A., Deshpande I., Wadasadawala T., Ray P., Sen S, & De A. (2021). EpCAM-Mediated Cellular Plasticity Promotes Radiation Resistance and Metastasis in Breast Cancer. Frontiers in Cell and Developmental Biology, 8, 597673.
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