Go Back   Science Forums Biology Forum Molecular Biology Forum Physics Chemistry Forum > Molecular Research Topics Forum > Protocols and Methods Forum
Register Search Today's Posts Mark Forums Read

Protocols and Methods Forum Post Any Protocol, Method, Technique, Procedure or Tips / Troubleshooting for any Molecular Biology Technique.


Silver staining

Silver staining - Protocols and Methods Forum

Silver staining - Post Any Protocol, Method, Technique, Procedure or Tips / Troubleshooting for any Molecular Biology Technique.


Reply
 
LinkBack Thread Tools Display Modes
  #1  
Old 08-19-2005, 01:18 PM
Chris B.
Guest
 
Posts: n/a
Default Silver staining



Hello,
I need to stain polylisine - free and conjugated with peptide. Coomasie
Brillant Blue fails with free polylysine and - for my suprise - so does
silver.
I'm using silver staining method from Amersham Technical Manual (as it say -
based on Heukeshoven and Dernick 1985) but they do not describe which
aminoacids are essential for detection.
Any help?
Regards,
Chris B.


Reply With Quote
  #2  
Old 08-28-2005, 09:06 AM
heartthrob
Guest
 
Posts: n/a
Default Silver staining

hello,
Maybe, the using of this method is limitted by F.W. of your
polylisine. It cannot apply to a small peptide. You can change another
polylisine with greater F.W.
Good luck!

Reply With Quote
  #3  
Old 08-29-2005, 08:48 AM
Andreas Giessauf
Guest
 
Posts: n/a
Default Silver staining



Chris B. schrieb:
I used already Ag staining method described by Blum et al.

Electrophoresis 8 (1987) 93-99 for protein staining.

Also, this autor didn't describe which residues are necessary.

But there are just two possibilities how the method works:

(1) Ag+ from the fixing solution is reduced to Ag (colloid)

(2) Ag+ reacts with free -SH groups and forms an Ag-S-R complex

In the first case there must be amino acid residues present

that can be oxidized by Ag+ (cysteines and others).

In the second case there must be amino acid residues present

that can form sulfides with Ag+-salts (cysteines are the only

possibility).

Best regards,

Andreas Giessauf

Reply With Quote
  #4  
Old 08-30-2005, 01:24 PM
Dr Engelbert Buxbaum
Guest
 
Posts: n/a
Default Silver staining

Chris B. wrote:


What is the molecular weight of your peptides? If it is too small, they
will be eluted from the gel during the staining procedure. Cross-linking
with glutardialdehyde may help in this case.

Also, polylysine will have a considerable positive charge, which would
interfere with Ag+ binding. Staining with an amine reactive fluorescent
probe may be the preferred option, either before or after
electrophoresis. Check the Molecular Probes catalog ([Only registered users see links. ]) for
options.
Reply With Quote
Reply

Tags
silver , staining


Thread Tools
Display Modes

Posting Rules
You may not post new threads
You may not post replies
You may not post attachments
You may not edit your posts

BB code is On
Smilies are On
[IMG] code is On
HTML code is Off
Trackbacks are On
Pingbacks are On
Refbacks are On

Forum Jump

Similar Threads
Thread Thread Starter Forum Replies Last Post
Immidiate dissappearing of bands on DNA sequencing gels after silver staining himanshu SDS-PAGE Gel Electrophoresis Forum 2 07-24-2008 06:17 PM
Problems with silver staining terence diane fabella Protein Forum 2 01-16-2008 06:04 PM
USP silver staining method help leelavati shetty Protein Forum 0 01-16-2008 05:17 AM
Silver staining Chris B. Protein Forum 3 08-30-2005 01:24 PM
Silver staining Western membranes EK Protocols and Methods Forum 14 09-27-2003 02:37 PM


All times are GMT. The time now is 03:24 PM.


Powered by vBulletin® Version 3.8.4
Copyright ©2000 - 2014, Jelsoft Enterprises Ltd.
Copyright 2005 - 2012 Molecular Station | All Rights Reserved
Page generated in 0.13211 seconds with 16 queries