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Invitrogen
Multispecies ERK1/ERK2 In-Cell ELISA Kit, Colorimetric
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Multispecies ERK1/ERK2 In-Cell ELISA Kit, Colorimetric
Relative total and phospho-ERK1/2 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 100 ng/mL EGF for five minutes immediately prior to fixation. At 10µM U0126, the phosphorylation of ERK1/2 was inhibited to levels similar to that observed in HeLa cells that were not stimulated wit... View More
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Product Specifications
Sample type/volume
Supernatant
10,000 cells
Hands-on time
1 hr 20 min
Time-to-result
24 hr
Homogenous (no wash)
No
Instrument
Absorbance Microplate Reader
Product size
96 Tests
Contents
Blocking Buffer
20X Tris Buffered Saline
Surfact-Amps 20
Surfact-Amps X-100
HRP Conjugate
TMB Substrate
TMB Stop Solution
Janus Green Whole-Cell Stain
Elution Buffer
Thin Plate Seal Assembly
Components included only in the target-specific kits: Antibody #1 and Antibody #2
20X Tris Buffered Saline
Surfact-Amps 20
Surfact-Amps X-100
HRP Conjugate
TMB Substrate
TMB Stop Solution
Janus Green Whole-Cell Stain
Elution Buffer
Thin Plate Seal Assembly
Components included only in the target-specific kits: Antibody #1 and Antibody #2
Shipping conditions
Wet ice
Storage
2-8°C
Protein name
ERK1/2
Target site-specificity
Protein family
Other Proteins
Assay kit format
In-cell ELISA Kit
Detector antibody conjugate
HRP
Label or dye
HRP
About This Kit
The ERK1/2 Multispecies In-Cell ELISA Colorimetric Detection Kit is a simple method for quantifying intracellular proteins in whole cells.
Principle of the method
To perform the assay, cells are first plated, treated and fixed. Expression of the protein(s) of interest is monitored in wells of a microplate using target-specific primary antibodies (see the Important Product Information section for antibodies included in each kit) and a horseradish peroxidase (HRP)-conjugated detection reagent. The kit is supplied with a whole-cell stain to control for differences in cell plating, which is important when measuring relative levels of a protein with different treatments or assessing its post-translational modification (PTM) form. After staining, the results are analyzed by normalizing the absorbance (HRP activity) values to cell number, which adjusts for the cell plating differences among the wells.
Rigorous validation
Each manufactured lot of this ELISA kit is quality tested for criteria such as sensitivity, specificity, precision, and lot-to-lot consistency. See manual for more information on validation.
Target information
ERK1 and ERK2 are widely expressed and are involved in the regulation of meiosis, mitosis, and postmitotic functions in differentiated cells. Many different stimuli, including growth factors, cytokines, virus infection, ligands for heterotrimeric guanine nucleotide-binding protein (G protein)-coupled receptors and transforming agents, activate the ERK1 and ERK2 pathways. When growth factors bind to the receptor tyrosine kinase, Ras interacts with Raf, the serine/threonine protein kinase and activates it as well. Once actived, Raf phosphorylates serine residue in 2 further kinases, MEK1/2, which in turn phosphorylates tyrosine/threonine in extracellular-signal regulated kinase (ERK) 1/2. Upon activation, the ERKs either phosphorylate a number of cytoplasmic targets or migrate to the nucleus, where they phosphorylate and activate a number of transcription factors such as c-Fos and Elk-1.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
Performance Guarantee
If an Invitrogen™ immunoassay doesn't perform as described on our website or datasheet,we'll replace the product at no cost to you, or provide you with a credit for a future purchase.*
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