WB
Various whole cell lysates were separated by 4-20% SDS-PAGE, and the primary antibody was used at 4°C, 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.
IHC
Immunofluorescence analysis of A549. 1,primary Antibody(red) was diluted at 1:200(4°C overnight). 2, Goat Anti Rabbit IgG (H&L) - Alexa Fluor 594 Secondary antibody was diluted at 1:1000(room temperature, 50min).3, DAPI(blue) 10min.
IF-P
Immunofluorescence analysis of rat-spleen tissue. 1,PERK Antibody(red) was diluted at 1:200(4°C,overnight). 2, Cy3 labled Secondary antibody was diluted at 1:300(room temperature, 50min).3, Picture B: DAPI(blue) 10min. Picture A:Target. Picture B: DAPI. Picture C: merge of A+B.