Related topics: Alphabetical list, Cancer biology, Cell cycle, Signal transduction
See list of additional phospho-MAPK antibodies at Linscott’s Directory
On October 3, 2018
Michelle Pedersen wrote:
I have tested this antibody on mouse HepG2 cells, stimulated with 10nM insulin for 10 minutes.
The cells were lysed in kinexus lysis buffer (see below) and 50ug protein were loaded in each well for western blot.
The membrane was left to incubate over night at 4 C in blocking buffer (milk).
The antibody worked very well.
20 mM MOPS, pH 7.0 (any other buffer at this pH could be substituted);
• 2 mM EGTA (to bind calcium);
• 5 mM EDTA (to bind magnesium and manganese);
• 30 mM sodium fluoride (to inhibit protein-serine phosphatases);
• 60 mM β-glycerophosphate, pH 7.2 (to inhibit protein-serine phosphatases);
• 20 mM sodium pyrophosphate (to inhibit protein-serine phosphatases);
• 1 mM sodium orthovanadate (to inhibit protein-tyrosine phosphatases);
• 1% Triton X-100 (can be substituted with 1% Nonidet P-40) Important Note: Do not add if you intend to first prepare a cytosolic fraction.
• 1 mM phenylmethylsulfonylfluoride (to inhibit proteases);
• 3 mM benzamidine (to inhibit proteases);
• 5 µM pepstatin A (to inhibit proteases);
• 10 µM leupeptin (to inhibit proteases);
• 1 mM dithiothreitol (to reduce disulphide linkages)
adjust to pH 7.2
On July 13, 2016
Signe Petersen, Aarhus University wrote:
PHOSPHO-MAPK (ERK1/2) (THR202/TYR204) RABBIT MAB CELL SIGNALING #4370
I have used this antibody several times for western blotting of primary hippocampal neurons stimulated with BDNF. Blots have always been succesful. It detects two distinct bands at 44 and 42 kDa. There are very little or no unspecific bands to be detected.
On February 12, 2015
Dusan Matusica, Flinders University wrote:
I use this antibody routinely for western blotting of PC12 cell lysates or NSC34 cell lysates following growth factor stimulation (NGF/BDNF). It works brilliantly at 1:2000 on as little as 1µg of protein per lane, blocked with 3% BSA in TBST.
On January 29, 2015
Daryl Jones, Mayo Clinic wrote:
I used this for Western blotting, it worked well at a 1:2000 dilution on neuroblastoma cell lysates, following BDNF treatment of live cells
On September 19, 2014
Maria Lume, Helsinki University wrote:
Detects pERK levels (kDA 42,44) very nicely in WB (dilution 1:2000, 5%BSA/TBS/Tween).
Have tested it on lysate from MG87 cell-line as well as on embryonic mouse kidney total lysate (E14).
On February 9, 2014
Simon Glerup, Aarhus University wrote:
Excellent antibody for Western blotting.
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. Two intense band around 50 kDa were induced by BDNF.
WB dilution 1:1000
On January 29, 2014
Simon Mølgaard, Aarhus University wrote:
I have also tested this on cultured hippocampal neurons stimulated with 1 nM of BDNF for 10 minutes. This works great to detect phosphorylation of MAPK.
On December 5, 2012
Ditte Olsen, Aarhus University wrote:
I tested this antibody in 293 cell cultures which had been stimulated with 10nM insulin. It worked excellent in detecting insulin-induced phosphorylation of MAPK using Western blot. I diluted it 1:2000
On November 22, 2012
Simon Jensen, Aarhus University wrote:
This antibody was tested on 293 cells in culture. The cells were stimulated with either 0, 1, 10 or 100nM of insulin before being fixed and stained. This antibody showed high degree of staining compared to unstimulated. 10nM showed the highest staining. This antibody worked very well in detecting phosphorylation of Mapk. It was also tested on primary hippocampal neurons where it showed great difference between stimulated and unstimulated.
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