Alexa Fluor® 647 Mouse anti-EGF Receptor (pY845)
Clone 12A3 (RUO)
- Brand BD Phosflow™
- Vol. Per Test 20 µl
- Isotype Mouse IgG1, κ
- Reactivity Human (QC Testing) Mouse, Rat (Reported)
Intracellular staining (flow cytometry) (Routinely Tested)
- Immunogen Phosphorylated Human EGF Receptor Peptide
- Storage Buffer Aqueous buffered solution containing BSA and ≤0.09% sodium azide.
- Regulatory Status RUO
Regulatory Status Legend
Epidermal Growth Factor (EGF) elicits a variety of cellular responses that are initiated by EGF Receptor (EGFR) binding and activation of intrinsic tyrosine kinase activity. EGFR, also known as ErbB1 or HER1, is a member of the ErbB class of receptor protein tyrosine kinases. It has an extracellular ligand-binding domain, a single transmembrane region, and a cytoplasmic region containing a protein tyrosine kinase domain and a c-terminal regulatory domain with many phosphorylation sites. Following ligand binding, EGFR forms homodimers and heterodimers with ErbB2. Specific C-terminal tyrosine residues are then autophosphorylated and, in turn, bind to adaptor proteins, kinases, or protein tyrosine phosphatases. In addition, c-Src-dependent phosphorylations at other sites, such as tyrosine 845 (Y845) in the kinase domain, regulate the receptor's kinase activity. Inappropriate expression or mutations of EGFR and/or deregulation of its signaling pathways are associated with many types of cancer, making EGFR a promising target for cancer therapies.
The 12A3 monoclonal antibody recognizes the phosphorylated Y845 in the protein tyrosine kinase domain of human EGFR.
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).
- Alexa Fluor® 647 fluorochrome emission is collected at the same instrument settings as for allophycocyanin (APC).
- Alexa Fluor® is a registered trademark of Molecular Probes, Inc., Eugene, OR.
- 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.
- Source of all serum proteins is from USDA inspected abattoirs located in the United States.
- 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.
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- Please refer to www.bdbiosciences.com/pharmingen/protocols for technical protocols.