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S6 In-Cell ELISA Kits


Microplate assays to measure total and phospho-S6 in whole cells.

Pierce S6 In-Cell ELISA Kits

Thermo Scientific Pierce S6 In-Cell ELISA Kits are colorimetric and fluorescent microplate assays optimized to measure total and phosphorylated ribosomal S6 protein (phospho-S6) in whole cells.

These ELISA kits for whole cells include a mouse monoclonal anti-S6 antibody, rabbit monoclonal anti-phospho-(Ser235/236) S6 antibody, detection reagents and all buffers required for either colorimetric or near infrared detection. The S6 In-Cell ELISA Kits enable simultaneous detection of both total and phospho-(Ser235/236) S6.

Highlights:

  • Measures total and phosphorylated ribosomal S6 protein in whole cells using a 96- or 384-well microplates
  • Two reliable detection formats available:
    • colorimetric detection kit for use with standard microplate readers
    • near infrared fluorescent detection kit for use with IR imaging system
  • Whole-cell detection requires no cell lysis, protein extraction or special sample preparation
  • Provides better data and higher throughput than quantitative Western blotting
  • Less costly (one well vs. one lane) than Western blotting

Product Details:

Upon recovery from serum-starvation, phosphorylation of ribosomal S6 protein increases >3-fold in NIH3T3. Relative total and phospho-S6 levels were measured in NIH3T3 cells using the (A) S6 Colorimetric In-Cell ELISA Kit and the (B) S6 Near Infrared In-Cell ELISA Kit. Cells were serum-starved overnight and stimulated with 20% fetal bovine serum 5 minutes immediately prior to fixation.The level of S6 phosphorylation increased 3- to 8-fold (colorimetric detection and near IR detection, respectively) in response to serum treatment where as total S6 protein levels were not significantly affected. Note: Absolute absorbance values cannot be directly compared between phospho and total S6 experiments because they use antibodies with different sensitivities.

Overview of S6 and phosphorylation of S6:

The ribosomal S6 protein is a component of the small subunit of ribsomes (40S) and is involved in the regulation of translation. The activity of S6 is regulated, in part, by the downstream effectors of mTOR, the ribosomal protein S6 kinase (S6K). Phosphorylation of S6 by S6K is important in the regulation of protein synthesis, cell size, cell-cycle progression, glucose homeostasis and survival of newborns. S6K activation and phosphorylation of S6 are implicated in translation efficiency of mRNAs with a 5'-terminal oligopyrimidine tract (TOP mRNAs) in certain physiological conditions. S6 phosphorylation occurs on 5 serine residues (Ser235, Ser236, Ser240, Ser244 and Ser247) within the C-terminus of the protein and are evolutionarily conserved in higher eukaryotes.The S6 In-Cell ELISA Kit enables simultaneous detection of both total and phospho-(Ser235/236) S6.

Related Resources:

Overview and Menu of Target-Specific In-Cell ELISA Kits
Dylight Fluor Spectral Properties (Ex/Em)

Related Products:

ELISA reagent reservoirs and sealing tape
Secondary Antibodies and Conjugates For ELISAs and Western Blotting
DyLight Fluor Conjugated Secondary Antibodies and Biotin Binding Proteins

Notice:

This product is for In Vitro experimental use only. Not for resale without express authorization.


Ordering Information
 
Product # Description Pkg. Size Instructions MSDS CofA Price
62207 S6 Colorimetric In-Cell ELISA Kit
Formulation: Multi-component kit
Sufficient For: 96 microplate wells of overnight-cultured cells

Kit Contents:
Blocking Buffer, 50mL
20X Tris Buffered Saline, 50mL
Surfact-Amps 20 (10% Detergent), 10mL
Surfact-Amps X-100 (10% Detergent), 10mL
HRP Conjugate, 0.14mL
TMB Substrate, 58mL
TMB Stop Solution, 55mL
Janus Green Whole-Cell Stain, 50mL
Elution Buffer, 2 x 30mL
Thin Plate Seal Assembly, 8 each
Anti-Phospho S6 (Ser 235/236) Antibody
Anti-S6 Antibody
96-test kit Product Instructions for product #62207 S6 Colorimetric In-Cell ELISA Kit MSDS for product #62207 S6 Colorimetric In-Cell ELISA Kit $380.00

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62212 S6 Near Infrared In-Cell ELISA Kit
Formulation: Multi-component kit
Sufficient For: 96 microplate wells of overnight-cultured cells

Kit Contents:
Blocking Buffer, 100mL
20X Tris Buffered Saline, 50mL
Surfact-Amps 20 (10% Detergent), 10mL
Surfact-Amps X-100 (10% Detergent), 10mL
DyLight 680 Goat Anti-Mouse IgG (H+L), 55µL
DyLight 800 Goat Anti-Rabbit IgG (H+L), 55µL
96-test kit Product Instructions for product #62212 S6 Near Infrared In-Cell ELISA Kit MSDS for product #62212 S6 Near Infrared In-Cell ELISA Kit $380.00

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Instructions | MSDS | CofA
Product Instructions | MSDS | CofA  

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