WB analysis of various samples using GTX24194 DNA-PKcs (phospho Thr2609) antibody.
Lane 1 : Protein ladder
Lane 2 : DNA Pkc phosphor
Lane 3 : DNA Pkc
Lane 4 : BSA
Loading : 10 μg
Dilution : 1 μg/mL

WB analysis of various samples using GTX24194 DNA-PKcs (phospho Thr2609) antibody. Lane 1-5 are nuclear extracts. Lane 6-10 are whole cell lysates
Lane 1 and 6 : Untreated Fus1 whole cell lysates
Lane 2 and 7 : IR-treated Fus1 whole cell lysates
Lane 3 and 8 : IR and DNAPK inhibitor-treated Fus1 whole cell lysates
Lane 4 and 9 : Untreated M059J whole cell lysates
Lane 5 and 10 : IR-treated M059J whole cell lysates
Loading : 20 μg
Dilution : 1:1000

Western blot using GeneTex Affinity Purified anti-DNAPKcs antibody (GTX24194) shows detection of a 460 kDa band corresponding to human DNAPKcs in various preparations. Lane 1: Fus1 untreated Lane 2: Fus1 IR (20Gy, 4h) Lane 3: Fus1 DNAPK inhibitor + IR Lane 4: MO59J (DNAPK-) untreated Lane 5: MO59J IR Lane 6: Fus1 untreated Lane 7: Fus1 IR (20Gy, 4h) Lane 8: Fus1 DNAPK inhibitor + IR Lane 9: MO59J untreated Lane 10: MO59J IR. Lanes 1-5 are nuclear extract whereas lanes 6-10 are whole cell lysates. MO59J is a cell line that lacks DNA-PKcs. FUS1 is the matched cell line complemented with a chromosomal fragment containing the DNA-PKcs gene. Approximately 20 μg of lysate was run on SDS-PAGE and transferred onto nitrocellulose, followed by reaction with a 1:1,000 dilution of anti-DNAPKcs antibody. Detection occurred using a 1:5,000 dilution of HRP-labeled Goat anti-Rabbit IgG for 1 hour at room temperature. A chemiluminescence system was used for signal detection (Roche) using a 1 min exposure time.

The data was published in the journal PLoS One in 2011. PMID: 21912620