WB
Various whole cell lysates were separated by 4-20% SDS-PAGE, and the primary antibody was used at 4℃, over night with a 1:2500 dilution . The Dylight 800-conjugated Goat anti-Rabbit antibody was used to detect the antibody. Lane1: HL60 - Human promyelocytic leukemia cell Lane2: HUVEC - Human umbilical vein endothelial cell Lane3: Human CAF - Human cancer-associated fibroblast Lane4: HCT116 - Human colorectal carcinoma Lane5: A549 - Human lung carcinoma Lane6: Jurkat - Human T lymphocyte leukemia Lane7: HT22 - Mouse hippocampal neuronal Lane8: Thp-1 - Human monocytic leukemia Lane9: HK-2 - Human proximal tubular epithelial Lane10: PC-12 - Rat adrenal pheochromocytoma Lane11: 3T3-L1 - Mouse embryonic fibroblast Lane12: PC-3 - Human prostate adenocarcinoma Lane13: U-14 - Mouse cervical carcinoma Lane14: 293F - HEK293 derivative, adapted for suspension culture.
WB
Western blot analysis of lysates from 1) 4T1, 2) 293 cells, (Green) primary antibody was diluted at 1:1000, 4°over night, Dylight 800 secondary antibody(Immunoway:RS23920)was diluted at 1:10000, 37° 1hour. (Red) Actin β Monoclonal Antibody(5B7) antibody was diluted at 1:5000 as loading control, 4° over night,Dylight 680 secondary antibody was diluted at 1:10000, 37° 1hour.
WB
Western blot detection of Phosphorylation of H2A.X at Serine 139 in 3T3 or Hydroxyurea-treated 3T3 cell lysates using Phospho-Histone H2A.X (Ser139) mouse mAb (1:2000 diluted).Predicted band size:15KDa.
WB
Various whole cell lysates were separated by 4-20% SDS-PAGE, and the membrane was blotted with anti-Histone H2A.X (phospho Ser139) antibody. The HRP-conjugated Goat anti-Rabbit IgG(H + L) antibody was used to detect the antibody. Lane 1: HepG2 treated with UV for 30 minutes Lane 2: PC-12 Lane 3: NIH-3T3 Lane 4: HEK293.
ICC/IF
Immunofluorescent analysis of Phosphorylation of H2A.X at Serine 139 in 3T3 or Hydroxyurea-treated 3T3 cells using Phospho-Histone H2A.X.
IHC
Mouse testis was stained with anti-Histone H2A.X (phospho Ser139) rabbit antibody.