FlowCellect™ EGFR/STAT3 Pathway Activation Detection Kit

FlowCellect™ EGFR/STAT3 Pathway Activation Detection Kit

FlowCellect

25 Tests / Kit

Millipore’s FlowCellect™ EGFR/STAT3 Activation Detection kit includes two directly conjugated phospho specific antibodies: pEGFR(Tyr1173)-Alexa Fluor® 488 and pSTAT3 (Tyr705)-Alexa Fluor® 647, along with optimized fixation, permeabilization, wash, and assay buffers to provide researchers the ability to detect the activation of EGFR and STAT3 at the receptor and downstream signaling level.
Antibodies against phospho-EGFR and phospho-STAT3 are provided in the kit to measure the activation state of each protein. A431 cells were used as a model system to demonstrate EGF mediated activation of EGFR as well as its downstream transcription activator STAT3. The treated cells were compared to untreated cells to verify the detection of both activated proteins by flow cytometry. When performing a dual color stain, the majority of the treated cells are double positive indicating activation of STAT3 through EGFR.
Sufficient reagents are provided for 25 2-color samples. The kit includes all optimized fluorescent labeled antibodies and buffers necessary for cell preparation and analysis.

The Epidermal Growth Factor Receptor (EGFR) is one of the Receptor Tyrosine Kinases which can activate the JAK/STAT pathway, one of three major signaling pathways downstream of EGFR. EGFR belongs to ErbB receptor superfamily, a group of transmembrane glycoprotein receptors, consisting of four members: EGFR (ErbB-1), ErbB-2 (c-neu, HER2), ErbB-3 (HER3), and ErbB-4 (HER 4).

STAT3 belongs to a family of conserved signaling molecules consisting of an oligomerization domain, a DNA binding domain, and a Src homology 2 (SH2) domain. STAT activation can be achieved not only by EGFR downstream signals, but also through the activation of the IL-6 pathway.

Deregulation of EGFR/STAT3 signaling due to alterations affecting the expression or function of a number of pathway components has long been associated with numerous forms of cancer, such as lung adenocarcinomas and breast cancer. Due to the complexity of the pathway and its involvement in many malignancies, the EGFR/STAT3 pathway is studied by numerous researchers looking for insight into the mechanisms involved in the formation and progression of cancers.

Monitoring the activation of EGFR and STAT3 simultaneously allows researchers to profile EGF mediated STAT3 activation within a cell type of interest. Changes in these signaling profiles can be used to monitor the effect of a particular upstream treatment targeting the pathway.

Flow Cytometry

Human

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4-8°C for antibodies and buffers

FlowCellect™ EGFR/STAT3 Pathway Activation Detection Kit

Flow Cellect

inCyte

Monoclonal Antibody

Research

Lot to lot in-process testing on all kit components

• 1. 20X Anti-phospho-EGFR (Tyr1173) AlexaFluor® 488 Antibody: (Part No. CS204825) One vial containing 150 µL.
• 2. 20X Anti-pSTAT3 (Tyr705), Alexa Fluor® 647 Antibody (Part No. CS205007) One vial containing 150 µL antibody.
• 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. CS202125) One bottle containing 13 mL buffer.

Mouse

Fluorescent