The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Use a concentration of 1 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
1/1000 - 1/10000. Detects a band of approximately 30 kDa (predicted molecular weight: 30 kDa).
Use at 2-5 µg/mg of lysate.
May play an important role in controlling cell growth and proliferation through the selective translation of particular classes of mRNA.
Belongs to the ribosomal protein S6e family.
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.
IHC image of RPS6 staining in human skin formalin fixed paraffin embedded tissue section, performed on a Leica Bond system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab70227, 1µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.
For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.
Western blot - RPS6 antibody (ab70227)
All lanes : Anti-RPS6 antibody (ab70227) at 0.04 µg/ml
Lane 1 : Whole cell lysate from HeLa cells at 50 µg Lane 2 : Whole cell lysate from HeLa cells at 15 µg Lane 3 : Whole cell lysate from HeLa cells at 5 µg Lane 4 : Whole cell lysate from 293T cells at 50 µg Lane 5 : Whole cell lysate from NIH3T3 cells at 50 µg
Predicted band size: 30 kDa Observed band size: 30 kDa
Exposure time: 30 seconds
Immunoprecipitation - RPS6 antibody (ab70227)
Detection of Human RPS6 by Immunoprecipitation. Samples: A) Whole cell lysate from HeLa cells (1 mg/IP; 1/4 of IP loaded) using affinity purified rabbit anti-RPS6
antibody ab70227 at 3 µg/mg lysate for IP and at 0.04 µg/ml for subequent WB detection. Second lane represents IgG IP control.
Hagner PR et al. Ribosomal protein S6 is highly expressed in non-Hodgkin lymphoma and associates with mRNA containing a 5' terminal oligopyrimidine tract. Oncogene : (2010).
Read more (PubMed: 21102526) »