Whole-cell protein lysates of treated cells were extracted and fractionated using sodium dodecyl sulfate-polyacrylamide gel electrophoresis and blotted onto nitrocellulose as previously described 14 (link). Protein expression was determined using the Western Lightning Plus-ECL detection system (NEL104001EA, PerkinElmer, MA, USA). Primary antibodies used: γ-H2AX (ab2893, abcam), activated caspase3 (ab32042, abcam), CHK2 pThr 68 (2661, Cell Signaling), CHK2 (3440, Cell Signaling), ATM pSer 1981 (5883, Cell Signaling) and GAPDH (MAB374, Millipore). Secondary antibodies used: anti-rabbit IgG, HRP-linked antibody (7074, Cell Signaling) and anti-mouse IgG, HRP-linked antibody (7076, Cell Signaling).
Chk2 pthr 68
Manufactured by Cell Signaling Technology
Sourced in United States
CHK2 pThr 68 is a primary antibody that detects endogenous levels of Chk2 protein only when phosphorylated at threonine 68. This antibody is intended for research use only.
Automatically generated - may contain errors
Lab products found in correlation
Atm pser 1981, by Cell Signaling Technology (3 mentions)
Ab2893, by Abcam (1 mentions)
Ab32042, by Abcam (1 mentions)
Anti rabbit igg hrp linked antibody, by Cell Signaling Technology (1 mentions)
Mab374, by Merck Group (1 mentions)
β actin a2066, by Merck Group (1 mentions)
Ab5821, by Abcam (1 mentions)
Fluorescein isothiocyanate (fitc), by Jackson ImmunoResearch (1 mentions)
Img 124a, by Novus Biologicals (1 mentions)
H 300, by Merck Group (1 mentions)
Bu 33, by Merck Group (1 mentions)
Cyanine3 (cy3), by Jackson ImmunoResearch (1 mentions)
3 protocols using chk2 pthr 68
1
Western Blot Analysis of DNA Damage Response
2
Protein Profiling by Western Blot
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Cell lysates, SDS-PAGE and Western blot were performed as described in [5 (link)]. Antibodies used were: ATM p-Ser1981 #13050, Chk2 p-Thr68 #2661, Chk2 #2662, Chk1 p-Ser345 #2341, Chk1 #2360, KAP1 #5868, cleaved caspase 3 (Asp175) #9661 (Cell Signaling Technology, Danvers, MA, USA), KAP1 p-Ser473 #644602 (Biolegend, San Diego, CA, USA), ATM #AF1655 (R&D Systems Bio-Techne, Minneapolis, MN, USA), vinculin #V9131, β-actin #A2066 (Millipore Sigma, Burlington, MA, USA). Detection was performed by exposure to standard X-ray films or by a CCD camera system (UVITEC, Cambridge, UK).
3
Immunofluorescence Analysis of Cellular Markers
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Antibodies included fibrillarin 1∶1000 (Abcam ab18380 or ab5821), CENP-A 1∶500 (Abcam 13939 or Upstate 07-574), TRF2 1∶200 (Imgenex IMG-124A or Novus NB110-57130), Ki67 antigen 1∶500 (Novocastra Laboratories Ltd.), H2AX-p 1∶300 (Millipore 05-636 or Abcam ab2893), ATM-p Ser1981 1∶250 (Cell Signaling 5883), Chk2-p Thr68 1∶250 (Cell Signaling 2661), SMC2 1∶300 (Cell Signaling 5394), SMC4 1∶300 (Cell Signaling 5547), goat polyclonal to DDDDK (FLAG) tag 1∶500 (Abcam ab1257), UBF 1∶150 (Santa Cruz H-300), and anti-BrdU 1∶400 (Sigma clone BU-33). Primary antibodies were detected using anti-mouse, anti-rabbit, or anti-goat secondary antibodies conjugated to Alexa Fluor 488, 594, 647 (Molecular Probes), FITC, Cy3, or Cy5 (Jackson Immunoresearch, Inc.).
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