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Preparation And Storage
Recommended Assay Procedures
Human Jurkat cells treated with H2O2 are suggested as a positive control; however, other cell types or methods may also be used for detection of phosphorylated PLCγ1.
The Alexa Fluor® 488-conjugated format has been evaluated using a human model system. The unconjugated form of this antibody (Cat. No. 612464) has been shown to react with human, mouse and rat cell lysates in western blot.
Product Notices
- This reagent has been pre-diluted for use at the recommended Volume per Test. We typically use 1 × 10^6 cells in a 100-µl experimental sample (a test).
- Alexa Fluor® 488 fluorochrome emission is collected at the same instrument settings as for fluorescein isothiocyanate (FITC).
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- The Alexa Fluor®, Pacific Blue™, and Cascade Blue® dye antibody conjugates in this product are sold under license from Molecular Probes, Inc. for research use only, excluding use in combination with microarrays, or as analyte specific reagents. The Alexa Fluor® dyes (except for Alexa Fluor® 430), Pacific Blue™ dye, and Cascade Blue® dye are covered by pending and issued patents.
- Alexa Fluor® is a registered trademark of Molecular Probes, Inc., Eugene, OR.
- Caution: Sodium azide yields highly toxic hydrazoic acid under acidic conditions. Dilute azide compounds in running water before discarding to avoid accumulation of potentially explosive deposits in plumbing.
- Source of all serum proteins is from USDA inspected abattoirs located in the United States.
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
Companion Products
The Phospholipase C (PLC) isozymes hydrolyze phosphatidyl inositol biphosphate to inositol triphosphate and diacylglycerol. The former causes release of calcium from the endoplasmic reticulum, while the latter is an activator of Protein Kinase C. Within the PLC family, PLCγ is the only member that contains SH2 and SH3 domains. These domains enable it to interact with receptor tyrosine kinases and become enzymatically activated via phosphorylation. It exists as two isoforms: 1) PLCγ1, which is ubiquitously expressed, and 2) PLCγ2, found primarily in the lymphoid system. PLCγ is essential for growth factor-induced cell motility and mitogenesis. PLCγ1-null mice exhibit retarded embryonic growth and lethality in midgestation. In addition, PDGF stimulation leads to phosphorylation of PLCγ1 at Tyr 783 and activation of hydrolyzing activity. Overexpression of PLCγ is evident in several forms of cancer, and it has been identified as a key mediator of PDGF-dependent cellular transformation. Thus, regulation of PLCγ activity by growth factors is involved in cell growth and transformation.
The 27/PLC antibody recognizes PLCγ1 phosphorylated at Y783. The fluorochrome-conjugated formats have been evaluated using a human model system. However, the unconjugated form of this antibody (Cat. no. 612464) has been shown to react with human, mouse, and rat cell lysates by western blot.
Development References (3)
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Chen P, Murphy-Ullrich JE, Wells A. A role for gelsolin in actuating epidermal growth factor receptor-mediated cell motility. J Cell Biol. 1996; 134(3):689-698. (Biology). View Reference
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Obermeier A, Tinhofer I, Grunicke HH, Ullrich A. Transforming potentials of epidermal growth factor and nerve growth factor receptors inversely correlate with their phospholipase C gamma affinity and signal activation. EMBO J. 1996; 15:73-82. (Biology).
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Yu H, Fukami K, Itoh T, Takenawa T. Phosphorylation of phospholipase Cγ1 on tyrosine residue 783 by platelet-derived growth factor regulates reorganization of the cytoskeleton. Exp Cell Res. 1998; 243:113-122. (Biology).
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Global - Refer to manufacturer's instructions for use and related User Manuals and Technical data sheets before using this products as described
Comparisons, where applicable, are made against older BD Technology, manual methods or are general performance claims. Comparisons are not made against non-BD technologies, unless otherwise noted.
For Research Use Only. Not for use in diagnostic or therapeutic procedures.
