Alexa Fluor® 647 Mouse Anti-Human PLCγ1 (pY783)
Clone 27/PLC (RUO)
- Brand BD Phosflow™
- Vol. Per Test 20 µl
- Isotype Mouse IgG1
- Reactivity Human (QC Testing)
Intracellular staining (flow cytometry) (Routinely Tested)
- Immunogen Phosphorylated peptide including the tyrosine 783 of human PLCγ1
- Storage Buffer Aqueous buffered solution containing BSA and ≤0.09% sodium azide.
- Regulatory Status RUO
Regulatory Status Legend
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.
Alexa Fluor® 647 conjugates are highly photostable and remain fluorescent over a broad pH range. The excitation and emission maxima are nearly identical to those of APC. However, APC tends to be brighter while Alexa Fluor® 647 is more optimal for intracellular applications. This fluorochrome exhibits uncommon photostability, making it an ideal choice for use in fluorescence microscopy. Due to nearly identical excitation and emission properties but different spillover characteristics, APC and Alexa Fluor® 647 cannot be used simultaneously.
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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.
- 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/pharmingen/protocols 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.