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.
Western blot
Immunostaining
Not Rated
IP
ELISA
Not Rated
Flow cytometry
Not Rated
Luminex
Not Rated
On February 9, 2014Simon Glerup, Aarhus University wrote:
5
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 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.
On November 5, 2020 Tomoki Ishibashi wrote:
I have used this antibody for IHC on Drosophila embryo. Dilution: 1:1000
On November 28, 2017 Anders Dalby wrote:
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.
On February 9, 2014 Simon Glerup, Aarhus University wrote:
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
On October 8, 2013 Yuval Yarkoni wrote:
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.