Hi everyone,
Does anyone have a suggestion for anti-Src and anti-pY418 Src antibodies that work well in immunofluorescence on cultured neurons?
Thanks a lot!
Dear Stefanie!
You look at following article, please:
Article Microtubule-mediated Src Tyrosine Kinase Trafficking in Neur...
eptide Antibodies
Polyclonal anti-Aplysia Src1 and Src2 antibodies (Pacific Immunology, Ramona, CA) were raised against peptides derived from respective N-terminal regions: GEKGSSTKYLPDPFQG (amino acids [aa] 10–25) for the rabbit anti-Src1 antibody; SNTAGDASPSHRLAENG (aa 14–30) for the goat anti-Src2 antibody. The sequence around the autophosphorylation site ARVIKEDE(pY)EARVG (aa 399–412) was used for the rabbit anti-pSrc2 (activated Src2) antibody. The pSrc2 antibody was first purified against the nonphosphorylated and then the phosphorylated peptide. Affinity-purified antibody stock solutions were prepared at 1 mg/ml in phosphate-buffered saline (PBS)/0.01% NaN3. To reduce binding to unspecific targets in Aplysia tissue, antibodies were first absorbed two times against Aplysia CNS proteins separated by SDS-polyacrylamide gel electrophoresis (PAGE) and transferred onto polyvinylidene fluoride membranes. The rabbit anti-active human Src antibody (Src PY418) was purchased from BioSource International (Invitrogen). Peptide blocking control experiments for Western blotting and immunocytochemistry were carried out by preincubation of antibody stock solutions (1 mg/ml in PBS), with an equal volume of the cognate or a control peptide (5 mg/ml in PBS) for 2 h at RT before antibody dilutions were prepared as described below.
Figure 2.
https://www.thermofisher.com/antibody/product/Phospho-SRC-Tyr419-Antibody-Polyclonal/44-660G
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