This antibody gave a positive signal in the following Human Whole Cell lysates; HEK293, HEK293 (Nuclear) and U2OS and FFPE human liver tissue sections.
ab96488 is raised using the same immunogen as ab42439. Batches sold under ab96488 have passed QC testing in Western Blot. In our hands, this product did not work in ICC/IF. For further information please contact our Customer Services team.
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.
This antibody detects multiple bands in addition to the band of interest, reflecting low levels of expression of DNA2. siRNA knockdown experiments have demonstrated that this antibody detects a band at ~120 kDa corresponding to human DNA2 (please see image courtesy of Julien Duxin).
Western blot - Anti-DNA2 antibody (ab96488)Image courtesy of Julien Duxin and Sheila Stewart
All lanes : Anti-DNA2 antibody (ab96488)
Lane 1 : U2OS cells plus control shRNA Lane 2 : U2OS cells plus shDNA2-1 shRNA Lane 3 : U2OS cells plus shDNA2-2 shRNA
Secondary All lanes : Anti-rabbit at 1/3000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 120 kDa Observed band size: 120 kDa
Western blot showing detection of DNA2 by ab42439. A 120 kDa band corresponding to DNA2 was detected in lysates of U2OS cells treated with control shRNA (lane 1) but not in those from knockdown cells treated with two different shRNAs for DNA2 (lanes 2 and 3). Following pre-blocking for 30 mins in 5% milk, PBS-Tween, the membrane was incubated overnight at 4°C with ab42439 at a dilution of 1/1000 in 5% milk, PBS-Tween. The membrane was then incubated with the secondary antibody at 1/3000 dilution in 5% milk, PBS-Tween for 2 hours at RT, and washed 3 x 10 mins with PBS-Tween prior to ECL detection.
IHC image of ab96488 staining in human liver carcinoma formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab96488, 1µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.
For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.
Biehs R et al. DNA Double-Strand Break Resection Occurs during Non-homologous End Joining in G1 but Is Distinct from Resection during Homologous Recombination. Mol Cell65:671-684.e5 (2017).
Read more (PubMed: 28132842) »