On June 30, 2020Nicola Micali, Yale University wrote:
0
Great antibody for IF. Concentration indicated by manufacturer’s protocol.
For more information see Micali et al. Cell Reports 2020, “Variation of Human Neural Stem Cells Generating Organizer States In Vitro before Committing to Cortical Excitatory or Inhibitory Neuronal Fates”. https://www.sciencedirect.com/science/article/pii/S2211124720305489
On May 5, 2020Margie Sutton, University of Texas MD Anderson Cancer Center wrote:
This antibody worked well with lysate of murine cortical neurons. The neurons were lysed with Kinexus lysis buffer. Clear bands were observed after 1 min. exposure.
Diltution 1:1000
Western blot
Immunostaining
Not Rated
IP
Not Rated
ChIP
Not Rated
ELISA
Not Rated
Flow cytometry
Not Rated
Luminex
Not Rated
EM
Not Rated
On October 3, 2018Michelle Pedersen wrote:
5
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.
Dilution: 1:1000
The membrane was left to incubate over night at 4 C in blocking buffer (milk).
The antibody worked very well.
Lysis buffer:
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
Western blot
Immunostaining
Not Rated
IP
Not Rated
ChIP
Not Rated
ELISA
Not Rated
Flow cytometry
Not Rated
Luminex
Not Rated
EM
Not Rated
On July 13, 2016Signe Petersen, Aarhus University wrote:
5
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.
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.
Western blot
Immunostaining
Not Rated
IP
Not Rated
ChIP
Not Rated
ELISA
Not Rated
Flow cytometry
Not Rated
Luminex
Not Rated
EM
Not Rated
On January 29, 2015Daryl Jones, Mayo Clinic wrote:
4
I used this for Western blotting, it worked well at a 1:2000 dilution on neuroblastoma cell lysates, following BDNF treatment of live cells
Western blot
Immunostaining
Not Rated
IP
Not Rated
ChIP
Not Rated
ELISA
Not Rated
Flow cytometry
Not Rated
Luminex
Not Rated
EM
Not Rated
On September 19, 2014Maria Lume, Helsinki University wrote:
5
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).
Western blot
Immunostaining
Not Rated
IP
Not Rated
ChIP
Not Rated
ELISA
Not Rated
Flow cytometry
Not Rated
Luminex
Not Rated
EM
Not Rated
On February 9, 2014Simon Glerup, Aarhus University wrote:
5
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
Western blot
Immunostaining
Not Rated
IP
Not Rated
ChIP
Not Rated
ELISA
Not Rated
Flow cytometry
Not Rated
Luminex
Not Rated
EM
Not Rated
On January 29, 2014Simon Mølgaard, Aarhus University wrote:
5
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.
Dilution 1:200
Western blot
Not Rated
Immunostaining
IP
Not Rated
ChIP
Not Rated
ELISA
Not Rated
Flow cytometry
Not Rated
Luminex
Not Rated
EM
Not Rated
On December 5, 2012Ditte Olsen, Aarhus University wrote:
5
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
Western blot
Immunostaining
Not Rated
IP
Not Rated
ChIP
Not Rated
ELISA
Not Rated
Flow cytometry
Not Rated
Luminex
Not Rated
EM
Not Rated
On November 22, 2012Simon Jensen, Aarhus University wrote:
5
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.
Dilution 1:200
On June 30, 2020 Nicola Micali, Yale University wrote:
Great antibody for IF. Concentration indicated by manufacturer’s protocol.
For more information see Micali et al. Cell Reports 2020, “Variation of Human Neural Stem Cells Generating Organizer States In Vitro before Committing to Cortical Excitatory or Inhibitory Neuronal Fates”.
https://www.sciencedirect.com/science/article/pii/S2211124720305489
On May 5, 2020 Margie Sutton, University of Texas MD Anderson Cancer Center wrote:
WB dilution 1:2000
IF rating: 5 stars
IHC rating: 4 stars
For more information see Sutton et al. Cell Reports 2019, “DIRAS3 (ARHI) Blocks RAS/MAPK Signaling by Binding Directly to RAS and Disrupting RAS Clusters”
https://www.sciencedirect.com/science/article/pii/S221112471931527X#sec5
On July 1, 2019 Trine Rasmussen wrote:
This antibody worked well with lysate of murine cortical neurons. The neurons were lysed with Kinexus lysis buffer. Clear bands were observed after 1 min. exposure.
Diltution 1:1000
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.
Dilution: 1:1000
The membrane was left to incubate over night at 4 C in blocking buffer (milk).
The antibody worked very well.
Lysis buffer:
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.
Dilution 1:200
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.
Dilution 1:200