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I started to learn western blot from Denatured protein technique.
What is the purpose to run sample with native protein ?
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02 Nov 2011 - 01:39 -
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Topics I've Started
What is different between Native protein and Denatured protein samples.
08 May 2013 - 05:01 AM
Is there any way to get new topic on the forum on my smart phone?
04 April 2013 - 11:53 PM
I just would like to get notification on my phone when someone post new question ..
I guessed that I can use news feed or RSS ... but I don't know how to get one ?
or is there any application ( I use iPhone ) that can link to this forum ?
I guessed that I can use news feed or RSS ... but I don't know how to get one ?
or is there any application ( I use iPhone ) that can link to this forum ?
stimulate vs activate
12 November 2012 - 02:36 AM
Hello
Which one should I use in term of recepter and ligand.
Ligand activate recepter
Licand stimulate recepter
Which one should I use in term of recepter and ligand.
Ligand activate recepter
Licand stimulate recepter
Laemmli buffer a bit green or yellow after boiling!!?!?
24 October 2012 - 01:12 AM
Bromophenol blue
pH =3.0 - 4.6; color change = yellow to blue-violet
After i boiling my loading sample, it was looked like green-yellow. Probably the pH was between 3 - 3.5 I guess.
What could be the reason on my Laemmli buffer ? to make more acidic to my loading sample?
Does low pH could be the problem like on my images?
my laemmli buffer
Laemmli buffer for 10 mL
dd H2O - 3.29mL
0.5 M Tris-HCl pH 6.8 - 1.25 mL
Glycerol - 1mL
10% SDS - 2 mL
1% Bromphenolblue - 0,5mL
0.5 M EDTA - 0.005 mL
(Roche complete protease inhibitor 1 tablet and dissolve in water 2mL) - 1 mL
50 mM PMSF - 0.4mL
2-Mercap. 0.5mL
if you see on the picture, it look like spot
It was run on NuPage invitrogel 4-12% 1mm and MOPS running buffer from invitrogen as well.
pH =3.0 - 4.6; color change = yellow to blue-violet
After i boiling my loading sample, it was looked like green-yellow. Probably the pH was between 3 - 3.5 I guess.
What could be the reason on my Laemmli buffer ? to make more acidic to my loading sample?
Does low pH could be the problem like on my images?
my laemmli buffer
Quote
Laemmli buffer for 10 mL
dd H2O - 3.29mL
0.5 M Tris-HCl pH 6.8 - 1.25 mL
Glycerol - 1mL
10% SDS - 2 mL
1% Bromphenolblue - 0,5mL
0.5 M EDTA - 0.005 mL
(Roche complete protease inhibitor 1 tablet and dissolve in water 2mL) - 1 mL
50 mM PMSF - 0.4mL
2-Mercap. 0.5mL
It was run on NuPage invitrogel 4-12% 1mm and MOPS running buffer from invitrogen as well.
How to prepare blood serum for western blot?
19 September 2012 - 01:41 AM
Hi!
I found a protocol from
http://www.protocol-...osts/32178.html
What is final concentration after mixed blood serum with Laemmli buffer?
I'm normally working with cell pellets to run WB and I got 1 mg/mL for final concentration to get good band for my protein.
I found a protocol from
http://www.protocol-...osts/32178.html
Quote
For human serum I obtained 77 mg/mL
What is final concentration after mixed blood serum with Laemmli buffer?
I'm normally working with cell pellets to run WB and I got 1 mg/mL for final concentration to get good band for my protein.
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