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BD Phosflow™ Alexa Fluor® 647 Mouse Anti-Human PLCγ1 (pY783)
Clone 27/PLC (RUO)
Analysis of PLCγ1 (pY783) in activated human T leukemia cells. Jurkat cells (ATCC TIB152) were either stimulated with 5 mM hydrogen peroxide for 15 minutes (shaded histogram) or unstimulated (open histogram). The cells were fixed (BD Cytofix™ Fixation Buffer, Cat. No. 554655) for 10 minutes, then permeabilized (BD Phosflow™ Perm Buffer III, Cat. No. 558050) on ice for at least 30 minutes, and then stained with Alexa Fluor® 647 anti-PLCγ1 (pY783). Flow cytometry was performed on a BD FACSCalibur™ flow cytometry system.
Analysis of PLCγ1 (pY783) in activated human T leukemia cells. Jurkat cells (ATCC TIB152) were either stimulated with 5 mM hydrogen peroxide for 15 minutes (shaded histogram) or unstimulated (open histogram). The cells were fixed (BD Cytofix™ Fixation Buffer, Cat. No. 554655) for 10 minutes, then permeabilized (BD Phosflow™ Perm Buffer III, Cat. No. 558050) on ice for at least 30 minutes, and then stained with Alexa Fluor® 647 anti-PLCγ1 (pY783). Flow cytometry was performed on a BD FACSCalibur™ flow cytometry system.
Analysis of PLCγ1 (pY783) in activated human T leukemia cells. Jurkat cells (ATCC TIB152) were either stimulated with 5 mM hydrogen peroxide for 15 minutes (shaded histogram) or unstimulated (open histogram). The cells were fixed (BD Cytofix™ Fixation Buffer, Cat. No. 554655) for 10 minutes, then permeabilized (BD Phosflow™ Perm Buffer III, Cat. No. 558050) on ice for at least 30 minutes, and then stained with Alexa Fluor® 647 anti-PLCγ1 (pY783). Flow cytometry was performed on a BD FACSCalibur™ flow cytometry system.
Analysis of PLCγ1 (pY783) in activated human T leukemia cells. Jurkat cells (ATCC TIB152) were either stimulated with 5 mM hydrogen peroxide for 15 minutes (shaded histogram) or unstimulated (open histogram). The cells were fixed (BD Cytofix™ Fixation Buffer, Cat. No. 554655) for 10 minutes, then permeabilized (BD Phosflow™ Perm Buffer III, Cat. No. 558050) on ice for at least 30 minutes, and then stained with Alexa Fluor® 647 anti-PLCγ1 (pY783). Flow cytometry was performed on a BD FACSCalibur™ flow cytometry system.
Product Details
BD Phosflow™
Human (QC Testing)
Mouse IgG1
Phosphorylated peptide including the tyrosine 783 of human PLCγ1
Intracellular staining (flow cytometry) (Routinely Tested)
20 µl
AB_396921
Aqueous buffered solution containing BSA and ≤0.09% sodium azide.
RUO
Preparation And Storage
Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze. The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography. The antibody was conjugated to Alexa Fluor® 647 under optimum conditions, and unreacted Alexa Fluor® 647 was removed.
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).
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- Alexa Fluor® 647 fluorochrome emission is collected at the same instrument settings as for allophycocyanin (APC).
- 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.
Companion Products
Perm Buffer III RUO
Size
125 ML
Cat No.
558050
Fixation Buffer RUO
Size
100 ML
Cat No.
554655
557883 Rev. 3
Antibody Details
27/PLC
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.
557883 Rev. 3
Format Details
Alexa Fluor™ 647
Alexa Fluor™ 647 Dye is part of the BD red family of dyes. This is a small organic fluorochrome with an excitation maximum (Ex Max) at 653-nm and an emission maximum (Em Max) at 669-nm. Alexa Fluor™ 647 is designed to be excited by the Red laser (627-640 nm) and detected using an optical filter centered near 670-nm (e.g., a 660/20 nm bandpass filter). Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
Alexa Fluor™ 647
653 nm
669 nm
557883 Rev.3
Citations & References
View product citations for antibody "557883" on CiteAb
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).
557883 Rev. 3
Please refer to Support Documents for Quality Certificates
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.
Please refer to Support Documents for Quality Certificates
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.