ATM antibody | knockout validation | Abcam ab32420

This is a knockout-validated antibody summary, based on the publication "Evaluation of ATM heterozygous mutations underlying individual differences in radiosensitivity using genome editing in human cultured cells", as cited below [1]. Labome curates formal publications to compile a list of antibodies with unambiguous specificity within Validated Antibody Database (VAD).

ATM antibody | knockout validation | Abcam ab32420 figure 1
Figure 1. Western blotting analysis data showing the expression levels of ATM protein in primary fibroblasts derived from the six members of an A-T-affected family. The GAPDH antibody was used as a loading control. From [1].
Antibody information

Rabbit monoclonal IgG

Company: Abcam

Antibody: ATM

Catalog number: ab32420

Summary: Rabbit monoclonal IgG against a synthetic peptide within human ATM. Reacts with human by western blot, immunohistochemistry (paraffin), immunocytochemistry/immunofluorescence and flow cytometry. Unsuitable for immunoprecipitation.

Validation Method

Western blot

Sample

Human skin fibroblasts from ataxia-telangiectasia patients (heterozygous ATM null mutations, heterozygous carriers and normal individuals). Cells were lysed in RIPA buffer [2].

Blocking agent

Automated capillary-based western blotting system (WES, Proteinsimple).

Primary incubation

Automated capillary-based western blotting system (WES, Proteinsimple).

Secondary incubation

Automated capillary-based western blotting system (WES, Proteinsimple).

Detection

Automated capillary-based western blotting system (WES, Proteinsimple).

Clone note

The same clone (Y170) is sold as Abcam ab32420, ab216617; EMD Millipore 04-200.

References
  1. Royba E, Miyamoto T, Natsuko Akutsu S, Hosoba K, Tauchi H, Kudo Y, et al. Evaluation of ATM heterozygous mutations underlying individual differences in radiosensitivity using genome editing in human cultured cells. Sci Rep. 2017;7:5996 pubmed publisher
  2. 50mM Tris-HCl, pH 8.0, 150mM NaCl, 1% NP-40 (v/v), 0.1% sodium dodecyl sulphate, 0.5% sodium deoxycholate, 0.5mM SF, 2ng/ml pepstatin A, 10mg/ml leupeptin, 5mg/ml aprotinin.