Cell Signaling Technology

Product Pathways - PathScan ELISA

PathScan® Total S6 Ribosomal Protein Chemiluminescent Sandwich ELISA Kit #7337

When ordering five or more kits, please contact us for processing time and pricing at sales@cellsignal.com.

Kit Includes Volume Solution Color
S6 Ribosomal Protein Mouse Ab Coated Microwells 96 tests
S6 Ribosomal Protein Rabbit Detection Antibody 5.5 ml Green
Anti-rabbit IgG, HRP-Linked Ab 5.5 ml Red
Luminol/Enhancer Solution 3 ml Colorless
Stable Peroxide Buffer 3 ml Colorless
Sealing Tape 2 sheets
ELISA Wash Buffer (20X) 25 ml Colorless
ELISA Sample Diluent 25 ml Blue
Cell Lysis Buffer (10X) #9803 15 ml Yellowish

Note: 12 8-well modules –Each module is designed to break apart for 8 tests.
Note: Kit should be stored at 4°C with the exception of Cell Lysis Buffer, which is stored at –20°C (packaged separately).

Species Cross-Reactivity

H M

Reactivity Key:  H=Human  M=Mouse
Species enclosed in parentheses are predicted to react based on 100% sequence homology.

Description

The PathScan® Total S6 Ribosomal Protein Chemiluminescent Sandwich ELISA Kit is a solid phase sandwich enzyme-linked immunosorbent assay (ELISA) that detects endogenous levels of total S6 ribosomal protein with a chemiluminescent readout. Chemiluminescence ELISAs often have a wider dynamic range and higher sensitivity than conventional chromogenic detection. This chemiluminescent ELISA, which is offered in low volume microplates, shows increased signal and sensitivity while using smaller sample size. A S6 Ribosomal Protein Mouse mAb has been coated on the microwells. After incubation with cell lysates, the S6 ribosomal protein is captured by the coated antibody. Following extensive washing, S6 Ribosomal Protein Rabbit Antibody is added to detect the captured total S6 ribosomal protein. Anti-rabbit IgG, HRP-linked Antibody is then used to recognize the bound detection antibody. Chemiluminescent reagent is added for signal development. The magnitude of light emission, measured in relative light units (RLU), is proportional to the quantity of totalS6 ribosomal protein.Antibodies in kit are custom formulations specific to kit.

Specificity / Sensitivity

PathScan® Total S6 Ribosomal Protein Chemiluminescent Sandwich ELISA Kit #7337 detects endogenous levels of total S6 ribosomal protein in human and mouse cells. This kit detects proteins from the indicated species, as determined through in-house testing, but may also detect homologous proteins from other species.

Sensitivity

Sensitivity

Figure 1. Relationship between protein concentration from nonphospho cell lysate and PDGF-treated NIH/3T3 cell lysate and immediate light generation with chemiluminescent substrate is shown. NIH/3T3 cells (80% confluence) were lysed without the addition of phosphatase inhibitors to the lysis buffer (nonphospho lysate) or treated with hPDGF-BB #8912 (50 ng/ml, 30 min at 37ºC) and lysed with Cell Lysis Buffer (10X) #9803. Graph inset corresponding to the shaded area shows high sensitivity and a linear response at the low protein concentration range.

Background

One way that growth factors and mitogens effectively promote sustained cell growth and proliferation is by upregulating mRNA translation (1,2). Growth factors and mitogens induce the activation of p70 S6 kinase and the subsequent phosphorylation of the S6 ribosomal protein. Phosphorylation of S6 ribosomal protein correlates with an increase in translation of mRNA transcripts that contain an oligopyrimidine tract in their 5' untranslated regions (2). These particular mRNA transcripts (5'TOP) encode proteins involved in cell cycle progression, as well as ribosomal proteins and elongation factors necessary for translation (2,3). Important S6 ribosomal protein phosphorylation sites include several residues (Ser235, Ser236, Ser240, and Ser244) located within a small, carboxy-terminal region of the S6 protein (4,5).

  1. Dufner, A. and Thomas, G. (1999) Exp. Cell Res. 253, 100-109.
  2. Peterson, R.T. and Schreiber, S.L. (1998) Curr. Biol. 8, R248-R250.
  3. Jefferies, H.B. et al. (1997) EMBO J. 16, 3693-3704.
  4. Ferrari, S. et al. (1991) J. Biol. Chem. 266, 22770-22775.
  5. Flotow, H. and Thomas, G. (1992) J. Biol. Chem. 267, 3074-3078.

Application References

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Protocols

* Product-specific protocol.

Companion Products

For Research Use Only. Not For Use In Diagnostic Procedures.

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