Alexa Fluor® 647 Mouse anti-p130Cas (pY249)
Clone J169-757.12.2 (RUO)
- Brand BD Phosflow™
- Vol. Per Test 20 µl
- Isotype Mouse IgG2b, κ
- Reactivity Human (QC Testing)
Intracellular staining (flow cytometry) (Routinely Tested)
- Immunogen Phosphorylated Human p130Cas
- Storage Buffer Aqueous buffered solution containing BSA and ≤0.09% sodium azide.
- Regulatory Status RUO
Regulatory Status Legend
p47v-crk (v-Crk) is the product of a transforming gene, v-crk, that was isolated from avian sarcoma viruses. The v-Crk protein is a fusion product of viral Gag protein and a part of cellular Crk that includes SH2 and SH3 domains. v-Crk-induced transformation increases tyrosine phosphorylation of several cellular proteins, including p130Cas (CRK-associated substrate). The p130Cas is tightly associated with v-Crk via the SH2 domain of v-Crk. Tyrosine phosphorylation of p130Cas occurs in conjunction with cellular transformation in cells that express v-Src or v-Crk. This phosphorylation leads to a change in p130Cas localization from the cytoplasm to the cell membrane and, possibly, to the nucleus. Since p130Cas also associates with v-Src, it may be a v-Src substrate. Several phosphorylation sites have been described in p130Cas upon Fibroblast Growth Factor stimulation, and phosphorylated tyrosine (Y249) might function as a binding site for the Crk-adaptor molecule.
The J169-757.12.2 monoclonal antibody recognizes the phosphorylated Y249 of human p130Cas. The orthologous phosphorylation sites in mouse and rat p130Cas are Y253 and Y347, respectively.
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).
- 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.
- 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® 647 fluorochrome emission is collected at the same instrument settings as for allophycocyanin (APC).
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- Alexa Fluor® is a registered trademark of Molecular Probes, Inc., Eugene, OR.
- Please refer to www.bdbiosciences.com/pharmingen/protocols for technical protocols.