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Ordering Information

  • : FCCS025106
  • : 25 Tests / Kit
  • Product Family Information

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Flow Cytometry (representative lot data): Dual Parameter Analysis of Total and Phospho ERK1/2 on Jurkat Cells As illustrated in dot plots A and B, un...

MAPK Signaling Pathway. As indicated in the diagram above, the MAPK signaling pathway has many branching points which regulate a large number of cell...

FlowCellect™ MAPK Activation Dual Detection Kit

Description:
FlowCellect™ MAPK Activation Dual Detection Kit
Trade Name:
FlowCellect
Qty/Pk:
25 Tests / Kit
Product Overview:
Millipore’s FlowCellect™ MAPK Activation Dual Detection kit includes two directly conjugated antibodies, a phospho-specific Anti-phospho-ERK1/2 (Thr202/Tyr204, Thr185/Tyr187)-PE and an Anti-ERK1/2-Alexa Fluor® 647 conjugated antibody to measure total levels of ERK. This two color flow cytometry kit is designed to measure the extent of MAPK phosphorylation relative to the total MAPK expression in any given cell population. By doing such, the levels of both the total and phosphorylated protein can be measured simultaneously in the same cell, resulting in a normalized and accurate measurement of MAPK activation after stimulation. Moreover, simultaneous measurement of both total and phospho-ERK1/2 confirms target specificity of the phosphorylation event. Together, a total and phospho antibody duo performed in multiplex provides an enhanced and more reliable detection of the phospho:total ratio within a mixed population.
All FlowCellect kits are supplied with optimized fixation, permeabilization, wash, and assay buffers to provide researchers the ability to detect the activation of ERK in flow applications. Sufficient reagents are provided for 25 two-color samples. The kit includes all optimized fluorescently labeled antibodies and buffers necessary for cell preparation and analysis.
Background Information:
Erk (Extracellular signal-Related Kinase) is a family of two, highly homologous proteins denoted as Erk1 (p44, MAPK3) and Erk2 (p42, MAPK1) that both function in the same pathway. The two proteins are often referred to collectively as Erk1/2 or p44/p42 MAP kinase. The Erk pathway is considered the classical, canonical MAPK (Mitogen-Activated Protein Kinase) signaling pathway. It is an evolutionarily conserved pathway that controls and is a critical regulator of the growth and survival through the promotion of cell proliferation and the prevention of apoptosis. Erk is involved in the control of many fundamental cellular processes including cell proliferation, survival, differentiation, apoptosis, motility and metabolism. Erk is activated by growth factor stimulation of receptor tyrosine kinases (RTKs), GPCR, and/or integrin stimulation. This activates the Ras-Raf-MEK-Erk pathway that results in the phosphorylation/activation of Erk1/2 (p44/p42) on the TxY motif (Thr202/Tyr204 and Thr185/Tyr187 for Erk1 & Erk2, respectively).

The regulation of a large number of cellular processes (such as cell proliferation/differentiation, cell survival and apoptosis) is dependent on the activation of MAPK signaling pathway, which can be measured by the phosphorylation of ERK1/2.
Key Applications:
Flow Cytometry
Species Reactivity:
Human
Usage Statement:
  • Alexa Fluor® is a registered trademark of Molecular Probes, Inc.
  • Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.
Quality Assurance:
Lot to lot in-process testing on all kit components
Components:
  • 1. 20X Anti-phospho-ERK1/2 (Thr202/Tyr204, Thr185/Tyr187) PE Conjugated Antibody: (Part No. CS203329) One vial containing 150 µL.
  • 2. 20X Anti-ERK1/2-Alexa Fluor® 647 Conjugated Antibody: (Part No. CS205075) One vial containing 150 µL.
  • 3. Fixation Buffer: (Part No. CS202122) One bottle containing 13 mL.
  • 4. 10X Wash Buffer: (Part No. CS202123) One bottle containing 13 mL.
  • 5. 5X Assay Buffer: (Part No. CS202124) One 55 mL bottle
  • 6. 1X Permeabilization Buffer: (Part No. CS203284) One bottle containing 13 mL.
Host:
Mouse
Storage Conditions:
4-8°C for antibodies and buffers
Detection Methods:
Fluorescent
Product Name:
FlowCellect™ MAPK Activation Dual Detection Kit
Alternate Names:
Flow Cellect
Flow Cytometry Software Type:
ExpressPro
Antibody Type:
Monoclonal Antibody
Flow Cytometry Market:
Research