Phospho-Tyr, merckmillipore, 05-321

I have used this antibody for IHC on Drosophila embryo. Dilution: 1:1000

I have used this antibody for both WB and IP and it works very well in both categories.

Phosphorus-32 labelled primary murine neurons were lysed and by using 1ug/mL i was able to IP and detect clear bands corresponding to tyrosine-phosphorylated proteins.

For WB i use 1:1000 dilution. Works well but with little background.

We stimulated SY5Y neuroblastoma cells transfected with human TrkB with increasing concentrations of BDNF for 15 min. Cells were lysed in TNE buffer containing protease and phosphatase inhibitors. Lysates were analyzed by Western blotting and blots were developed using ECL Plus.
We observed a dose-dependent response to BDNF using this antibody and several bands became visible upon BDNF stimulation compared to unstimulated cells and untransfected cells.
WB dilution 1:1000

I have tested this antibody by Western blotting following immunoprecipitation. This works very well!

Dilution 1:1000

I have used 4G10 from Millipore (http://www.millipore.com/catalogue/item/05-321)
for WB at 0.1ug/ml on lysates from B cell lymphomas and it worked great.

I have used a home-made conjugate of 4G10 to Alexa 488 for intracellular flow. Since there is allways phosphorylated Tyr proteins in cells, it served as a positive control for my intracellular staining. Worked great.

Works for the detection of phosphorylated tyrosine residues in ELISA

Excellent for detecting phosphorylation of receptor tyrosine kinases by Western blotting following immunoprecipitation.

Dilution 1:1000