S6 Ribosomal Protein, Total, BioAssay(TM) ELISA Kit

To effectively promote growth and cell division in a sustained manner, growth factors and mitogens must upregulate translation (1,2). Growth factors and mitogens induce the activation of p70 S6 kinase, which in turn phosphorylates the S6 ribosomal protein. Phosphorylation of S6 ribosomal protein correlates with an increase in translation, particularly of mRNAs with an oligopyrimidine tract in their 5' untranslated regions (2). This group of mRNAs (5'TOP) encodes proteins involved in cell cycle progression and proteins that are part of the translational machinery, such as ribosomal proteins and elongation factors (2,3). The main in vivo S6 ribosomal protein phosphorylation sites, including Ser235, Ser236, Ser240 and Ser244, are located within a small 19 amino acid region in the S6 carboxy terminus (4,5). Total S6 Ribosomal Protein Sandwich ELISA Antibody Pair is being offered as an alternative to our Total S6 Ribosomal Protein Sandwich ELISA Kit. Capture and Detection antibodies (100X stocks) and an HRP-conjugated secondary antibody (1000X stock) are supplied. Sufficient reagents are provided for performing 4 x 96 well ELISAs. The S6 Ribosomal Protein Capture Antibody is coated in PBS overnight in a 96 well microplate. After blocking, cell lysates are added, followed by S6 Ribosomal Protein Detection Antibody and HRP-conjugated secondary antibody. HRP substrate, TMB, is added for color development. The magnitude of the absorbance at 450 nm is proportional to the quantity of Total S6 Ribosomal protein. Kit Components: S6 Ribosomal Protein Capture Antibody (100X), 1x0.4ml S6 Ribosomal Protein Detection Antibody (100X), 1x0.4ml Anti-Rabbit IgG HRP-Linked Antibody (1000X), 1x0.04ml Reagents not supplied: Phosphate Buffered Saline (PBS-20X) Phosphate Buffered Saline with Tween -20 (PBST-20X) Cell Lysis Buffer (10X) TMB Substrate STOP Solution Blocking Buffer- PBS+0.05% Tween-20, 1% BSA 96 Well Microplates Microplate Reader Storage and Stability: S6 Ribosomal Protein Capture Antibody and S6 Ribosomal Protein Detection Antibody are stored at 4 degrees C. Anti-Rabbit IgG-HRP Linked Antibody is stored at -20 degrees C. Stable for 6 months For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.