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ERK1-ERK2 In-Cell ELISA Kits


Microplate assays to measure total and phospho-ERK1/2 in whole cells.

Pierce ERK1/2 In-Cell ELISA Kits

Thermo Scientific Pierce ERK1/2 In-Cell ELISA Kits are colorimetric and fluorescent microplate assays optimized to measure total and phosphorylated extracellular signal-regulated kinase 1 and 2 (phospho-ERK1/2) in whole cells.

These ELISA kits for whole cells complete kits include a mouse monoclonal anti-ERK1/2 antibody, rabbit monoclonal anti-phospho-(Thr202/Tyr204) ERK1/2 antibody, detection reagents and all buffers required for either colorimetric or near infrared detection. The ERK1/2 In-Cell ELISA Kits enable simultaneous detection of both total and phospho-(Thr202/Tyr204) ERK1/2.

Highlights:

  • Measures total and phosphorylated extracellular signal-regulated kinase 1 and 2 (ERK1/2) 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:

Phosphorylation of MAPK3/1 in HeLa Cells following treatment with EGF. Stimulation with EGF causes a 6-fold increase in the level of MAPK3/1 phosphorylation in HeLa cells. Relative total and phospho-MAPK3/1 levels were measured in HeLa cells using the ERK1/2 Colorimetric In-Cell ELISA Kit. Cells were serum-starved overnight and stimulated with 100ng/ml EFG for five minutes immediately prior to fixation.The level of MAPK3/1 phosphorylation increased approximately 6-fold in response to EGF stimulation where as total MAPK3/1 were not significantly affected. Note: Absolute absorbance values cannot be directly compared between phospho and total MAPK3/1 experiments because they use antibodies with different sensitivities.

Dose-response curve showing decrease in total and phospho-MAPK3/1 in HeLa cells following U0126 treatment Treatment with U0126 inhibits EGF-induced phosphorylation of MAPK3/1 in HeLa cells. Relative total and phospho-MAPK3/1 levels were measured in HeLa cells using the ERK1/2 Colorimetric In-Cell ELISA Kit. Cells were serum-starved overnight, treated with U0126 for two hours and then stimulated with 100ng/ml EFG for five minutes immediately prior to fixation. At 10µM U0126, the phosphorylation of MAPK3/1 was inhibited to levels similar to that observed in HeLa cells that were not stimulated with EGF (data not shown). Note: Absolute absorbance values cannot be directly compared between phospho and total MAPK3/1 experiments because they use antibodies with different sensitivities.

Overview of ERK1/2 and phosphorylation of ERK1/2:

ERK1 and ERK2 are also referred to as mitogen-activated protein kinase 3 and 1, respectively (MAPK3, MAPK1) and regulate a broad range of cellular processes including gene transcription, cytoskeletal organization, metabolic homeostasis, cell growth, and apoptosis. Within the MAPK/ERK signal transduction pathway, extracellular stimuli may be transmitted into the cell through a receptor tyrosine kinase (RTK) and relayed through MAPKK kinase phosphorylation of MAPK kinase which, in turn, phosphorylates MAPK family members. Dual phosphorylation of ERK1/2 on thr 202 and tyr 204 causes translocation into the cell nucleus and phosphorylation of transcriptional regulators. Disregulation of ERK1/2 is implicated in cancer phenotypes. The ERK1/2 In-Cell ELISA Kit enables simultaneous detection of both total and phospho-(Thr202/Tyr204) ERK1/2.

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
62206 ERK1/2 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 ERK 1/2 (Thr 202/Tyr 204) Antibody
Anti-ERK1/2 Antibody
96-test kit Product Instructions for product #62206 ERK1/2 Colorimetric In-Cell ELISA Kit MSDS for product #62206 ERK1/2 Colorimetric In-Cell ELISA Kit $380.00

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62211 ERK1/2 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
Thin Plate Seal Assembly, 8 each
Anti-Phospho ERK 1/2 (Thr 202/Tyr 204) Antibody
Anti-ERK1/2 Antibody
96-test kit Product Instructions for product #62211 ERK1/2 Near Infrared In-Cell ELISA Kit MSDS for product #62211 ERK1/2 Near Infrared In-Cell ELISA Kit $380.00

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Instructions | MSDS | CofA
Product Instructions | MSDS | CofA  

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