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Human/Mouse/Rat Phospho-MKK3 (S189) Cell-Based ELISA Kit

Human/Mouse/Rat Phospho-MKK3 (S189) Cell-Based ELISA Kit

Item no.	CBEL-MKK3-1
Manufacturer	Raybiotech
Amount	1 Plate Kit
Quantity options	1 Plate Kit 2 Plate Kit 5 Plate Kit
Category	Cell Biology Cell Status ELISA Senescence ELISA & Immunoassays ELISA Kits Immunoassay
Type	Elisa-Kit
Specific against	Human (Homo sapiens)
Host	Hamster - Armenian
ECLASS 10.1	32160605
ECLASS 11.0	32160605
UNSPSC	41116126
Shipping Condition	Cool pack
Available
Manufacturer - Category	Assay Kits\|Phosphorylation Assays\|ELISA Kits\|Cell-Based ELISA
Manufacturer - Targets	MKK3
Shipping Temperature	Blue ice
Storage Conditions	-20°C
Description	RayBio® Cell-Based Human/Mouse/Rat MKK3 (S189) Phosphorylation ELISA Kit. Suitable for adherent whole cell lines.
Compatible Sample Types	Adherent Cell Culture
Design Principle	Cell-based
Method of Detection	Colorimetric
Quantitative/Semi-Quantitative	Semi-Quantitative
Solid Support	96-well Microplate
Gene Symbols	MAP2K3, MEK3, MKK3, PRKMK3, SKK2
Kit Components	Cell Culture Microplate Wash Buffer A Wash Buffer B Fixing Solution HRP-Conjugated Secondary Antibody Blocking Buffer TMB One-Step Substrate Stop Solution Quenching Buffer
Other Materials Required	Distilled or deionized water 100 ml and 1 liter graduated cylinders Tubes to prepare sample dilutions Protease and Phosphatase inhibitors Precision pipettes to deliver 2 µl to 1 ml volumes Adjustable 1-25 ml pipettes for reagent preparation Benchtop rocker or shaker Microplate reader capable of measuring absorbance at 450 nm
Protocol Outline	Seed 10, 000-30, 000 cells into each well and incubate overnight. Apply various treatment, inhibitors or activators according to manufacture's instructions. Add 100 µl of Fixing Solution into each well and incubate for 20 min at RT with shaking. Add 200 µl of prepared 1 X Quenching Buffer and incubate 20 min at RT. Add 200 µl of Blocking Solution and incubate for 1 h at 37 °C. Add 50 µl of 1 X anti-phospho-protein specific antibody or anti-pan-protein specific antibody to each well and incubate for 2 h at RT. Add 50 µl of prepared 1 X HRP-Anti-Rabbit or Mouse IgG and incubate for 1 h at RT. Add 100 µl of TMB One-Step Substrate Reagent to each well. Incubate 30 min at RT. Add 50 µl of Stop Solution to each well. Read at 450 nm immediately.
UNSPSC Code	41116158
Protein Name & Synonyms	Dual specificity mitogen-activated protein kinase kinase 3 (MAP kinase kinase 3) (MAPKK 3) (EC 2.7.12.2) (MAPK/ERK kinase 3) (MEK 3) (Stress-activated protein kinase kinase 2) (SAPK kinase 2) (SAPKK-2) (SAPKK2)
Manufacturer - Specificity	The antibodies provided in this kit recognizes human/mouse/rat MKK3 phosphorylated at Serine-189 and total MKK3 for comparison.
Pathway	MAPK Signaling
Introduction	Protein phosphorylation is instrumental in the regulation of protein activity within a cell. It plays important roles in the living cells including proliferation, differentiation and metabolism. A large number of protein kinases and phosphatases have been extensively investigated, and have been shown to be involved in signal transduction pathways. The RayBio® Cell-Based Human/Mouse/Rat MKK3 (S189) ELISA kit is a very rapid, convenient and sensitive assay kit that can monitor the activation or function of important biological pathways in cells. It can be used for measuring the relative amount of MKK3 (S189) phosphorylation and screening the effects of various treatments, inhibitors (such as siRNA or chemicals), or activators in cultured human, mouse and rat cell lines. By determining MKK3 protein phosphorylation in your experimental model system, you can verify pathway activation in your cell lines without spending excess time and effort in preparing cell lysate and performing an analysis of Western Blot. In the Cell-Based MKK3 (S189) ELISA kit, cells are seeded into a 96 well tissue culture plate. The cells are fixed after various treatments, inhibitors or activators. After blocking, Anti-Phospho-MKK3 (S189) or Anti-MKK3 is pipetted into the wells and incubated. The wells are washed, and HRP-Conjugated anti-rabbit IgG and HRP-Conjugated anti-mouse IgG are added to the corresponding wells. The wells are washed again, a TMB substrate solution is added to the wells and color develops in proportion to the amount of protein. The Stop Solution changes the color from blue to yellow, and the intensity of the color is measured at 450 nm.

Note: The presented information and documents (Manual, Product Datasheet, Safety Datasheet and Certificate of Analysis) correspond to our latest update and should serve for orientational purpose only. We do not guarantee the topicality. We would kindly ask you to make a request for specific requirements, if necessary.

All products are intended for research use only (RUO). Not for human, veterinary or therapeutic use.

CBEL-MKK3-1.pdf

CBEL-MKK3-1-datasheet.pdf

CBEL-MKK3-1-safety-datasheet.pdf