The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
1/10000 - 1/200000. Detects a band of approximately 30 kDa (predicted molecular weight: 30 kDa).
Use at an assay dependent concentration.
1/100 - 1/250.
应用说明Is unsuitable for Flow Cyt.
功能May play an important role in controlling cell growth and proliferation through the selective translation of particular classes of mRNA.
序列相似性Belongs to the ribosomal protein S6e family.
翻译后修饰Ribosomal protein S6 is the major substrate of protein kinases in eukaryote ribosomes. The phosphorylation is stimulated by growth factors, tumor promoting agents, and mitogens. It is dephosphorylated at growth arrest.
ICC/IF image of ab80158 stained DU145 cells. The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody ab80158 at 1/200 dilution overnight at +4°C. The secondary antibody (green) was DyLight® 488 goat anti- rabbit (ab96899) IgG (H+L) used at a 1/250 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.
Western blot - Anti-RPS6 (phospho S235) antibody [EP1338(2)Y] - Ribosome Marker (ab80158)
All lanes : Anti-RPS6 (phospho S235) antibody [EP1338(2)Y] - Ribosome Marker (ab80158) at 1/1000 dilution
Lane 1 : HeLa (Human cervix adenocarcinoma epithelial cell) serum starved for 24 h. Whole cell lysates Lane 2 : HeLa (Human cervix adenocarcinoma epithelial cell) serum starved for 24 h, then 20% FBS incubated for 90 minutes. Whole cell lysates Lane 3 : HeLa (Human cervix adenocarcinoma epithelial cell) serum starved for 24 h, then 20% FBS incubated for 90 minutes. Whole cell lysates. Then the membrane was incubated with phosphatase.
Lysates/proteins at 15 µg per lane.
Secondary Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size : 30 kDa Observed band size : 30 kDa
Exposure time : 1 minute
Blocking and diluting buffer and concentration: 5% NFDM/TBST.