Human Phospho-MKK3 (S218/T222) DuoSet IC ELISA

Name: Human Phospho-MKK3 (S218/T222) DuoSet IC ELISA
Brand: R&D Systems
SKU: DYC5585-2

Catalog #: DYC5585-2 Datasheet / COA / SDS

Discontinued Product

DYC5585-2 has been discontinued.

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Summary Product Features Kit Content Other Reagents Required Product Datasheets Preparation and Storage Background

Human Phospho-MKK3 (S218/T222) DuoSet IC ELISA Summary

Assay Type

Solid Phase Sandwich ELISA

Format

96-well strip plate

Assay Length

4 hours 40 minutes (after plate preparation)

Sample Volume Required

Cell lysates (100 µL)

Assay Range

62.5 - 4,000 pg/mL

Sufficient Materials

Kits available for two, five, or fifteen 96-well plates*

Specificity

Please see the product datasheet

* Provided that the recommended microplates, buffers, diluents, substrates and solutions are used, and the assay is run as summarized in the Assay Procedure provided.

This DuoSet IC ELISA contains the basic components required for the development of sandwich ELISAs to measure phosphorylated in cell lysates. An immobilized capture antibody specific for binds both phosphorylated and unphosphorylated . After washing away unbound material, a biotinylated detection antibody is used to detect only phosphorylated receptor, utilizing a standard HRP format.

Product Features

Optimized capture and detection antibody pairings with recommended concentrations save lengthy development time
Development protocols are provided to guide further assay optimization
Assay can be customized to your specific needs
Available in 2, 5, and 15- (96-well) plate pack sizes
Economical alternative to Western blot

Kit Content

Capture Antibody
Conjugated Detection Antibody
Calibrated Immunoassay Standard or Control
Streptavidin-HRP

Other Reagents Required

PBS: (Catalog # DY006), or 137 mM NaCl, 2.7 mM KCl, 8.1 mM Na₂HPO₄, 1.5 mM KH₂O₄, pH 7.2 - 7.4, 0.2 µm filtered

Wash Buffer: (Catalog # WA126), or equivalent

Lysis Buffer*

IC Diluent*

Blocking Buffer*

Substrate Solution: 1:1 mixture of Color Reagent A (H₂O₂) and Color Reagent B (Tetramethylbenzidine) (Catalog # DY999)

Stop Solution: 2 N H₂SO₄ (Catalog # DY994)

Microplates: From Costar EIA Plate (Costar Catalog # 2592) or R&D Systems (Catalog # DY990), or equivalent

Plate Sealers: ELISA Plate Sealers (Catalog # DY992), or equivalent

*For the Lysis Buffer, IC Diluent, and Blocking BUffer recommended for a specific DuoSet ELISA Development Kit, please see the product

Product Datasheets

Product Datasheet

COA

SDS

Preparation and Storage

Shipping

The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below.

Stability & Storage

Store the unopened product at 2 - 8 °C. Do not use past expiration date.

Background: MKK3

Mitogen-activated protein kinase kinase 3 (MKK3), also known as MEK3, and the closely related MKK6 are activated by proinflammatory cytokines and environmental stresses. Activation occurs through phosphorylation by several upstream MAPK kinase kinases, including ASK1 and TAK1. Both MKK3 and MKK6 are dual specificity protein kinases, phosphorylating and activating the p38 MAP kinases at Thr and Tyr positions within the phosphoacceptor sequence Thr-Gly-Tyr.

Long Name:

Mitogen-activated Protein Kinase Kinase 3

Entrez Gene IDs:

5606 (Human); 26397 (Mouse); 303200 (Rat)

Alternate Names:

MAP kinase kinase 3; MAP2K3; MAPK/ERK kinase 3; MAPKK 3; MAPKK3PRKMK3dual specificity mitogen-activated protein kinase kinase 3; MEK3; MEK3MEK 3; mitogen activated protein kinase kinase 3; mitogen-activated protein kinase kinase 3; MKK3; MKK3EC 2.7.12.2; PRKMK3