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Dual Parameter Analysis of Total and Phospho PLC-g1 on A431Cells Unstimulated A431 cells are stained with both phospho-PLC-g1-Alexa Fluor 488 and Ant...
FlowCellect™ PLC-gamma 1 Activation Dual Detection Kit
Description:
FlowCellect™ PLC-gamma 1 Activation Dual Detection Kit
Trade Name:
FlowCellect
Qty/Pk:
25 Tests / Kit
Product Overview:
Millipore’s FlowCellect™ PLC-g1 Activation Dual Detection kit includes two directly conjugated antibodies, a phospho-specific Anti-phospho-PLC-g1 (Tyr837)-Alexa Fluor® 488 and an Anti-PLC-g1-PE conjugated antibody to measure total levels of PLC-g1. This two color flow cytometry kit is designed to detect the extent of PLC-g1 pathway activation by measuring PLC-g1 phosphorylation relative to the total PLC-g1 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 PLC-g1 activation after stimulation. Moreover, simultaneous measurement of both total and phospho-PLC-g1 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.
PLC-g1 is devoted to tyrosine kinase-dependent signaling for mitogenic responses to diverse agents such as growth factor activation of receptor tyrosine kinases and stimulation of immune cell receptors. Signaling pathways that depend on tyrosine kinases communicate with the PLC-g family. Because of its involvement in many key signaling cascades resulting from receptor tyrosine kinase activation, PLC-g1 plays a critical role in tumor progression and cancer-related illnesses. When stimulating A431 cells with EGF for 2 minutes the total protein level remains constant compared to unstimulated cells, while phospho-PLC-g1 levels are increased in all cells indicating that there is no competition between the two antibodies for their target epitopes.
PLC-g1 is devoted to tyrosine kinase-dependent signaling for mitogenic responses to diverse agents such as growth factor activation of receptor tyrosine kinases and stimulation of immune cell receptors. Signaling pathways that depend on tyrosine kinases communicate with the PLC-g family. Because of its involvement in many key signaling cascades resulting from receptor tyrosine kinase activation, PLC-g1 plays a critical role in tumor progression and cancer-related illnesses. When stimulating A431 cells with EGF for 2 minutes the total protein level remains constant compared to unstimulated cells, while phospho-PLC-g1 levels are increased in all cells indicating that there is no competition between the two antibodies for their target epitopes.
Background Information:
Millipore’s FlowCellect™ PLC-gamma 1 Activation Dual Detection kit includes two directly conjugated antibodies, a phospho-specific Anti-phospho-PLC-gamma 1 (Tyr837)-Alexa Fluor® 488 and an Anti-PLC-gamma 1-PE conjugated antibody to measure total levels of PLC-gamma 1. This two color flow cytometry kit is designed to detect the extent of PLC-gamma 1 pathway activation by measuring PLC-gamma 1 phosphorylation relative to the total PLC-gamma 1 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 PLC-gamma 1 activation after stimulation. Moreover, simultaneous measurement of both total and phospho-PLC-gamma 1 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.
PLC-gamma 1 is devoted to tyrosine kinase-dependent signaling for mitogenic responses to diverse agents such as growth factor activation of receptor tyrosine kinases and stimulation of immune cell receptors. Signaling pathways that depend on tyrosine kinases communicate with the PLC-gamma family. Because of its involvement in many key signaling cascades resulting from receptor tyrosine kinase activation, PLC-gamma 1 plays a critical role in tumor progression and cancer-related illnesses. When stimulating A431 cells with EGF for 2 minutes the total protein level remains constant compared to unstimulated cells, while phospho-PLC-gamma 1 levels are increased in all cells indicating that there is no competition between the two antibodies for their target epitopes.
PLC-gamma 1 is devoted to tyrosine kinase-dependent signaling for mitogenic responses to diverse agents such as growth factor activation of receptor tyrosine kinases and stimulation of immune cell receptors. Signaling pathways that depend on tyrosine kinases communicate with the PLC-gamma family. Because of its involvement in many key signaling cascades resulting from receptor tyrosine kinase activation, PLC-gamma 1 plays a critical role in tumor progression and cancer-related illnesses. When stimulating A431 cells with EGF for 2 minutes the total protein level remains constant compared to unstimulated cells, while phospho-PLC-gamma 1 levels are increased in all cells indicating that there is no competition between the two antibodies for their target epitopes.
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-PLC-g1 (Tyr783) Alexa Fluor® 488: (Part No. CS206377) One vial containing 150 µL.
- 2. 20X Anti-PLC-g1-PE: (Part No. CS206378) 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™ PLC-gamma 1 Activation Dual Detection Kit
Alternate Names:
Flow Cellect
Flow Cytometry Software Type:
ExpressPro & inCyte
Antibody Type:
Monoclonal Antibody
Flow Cytometry Market:
Research