Can anyone please suggest me a rapid silver staining protocol. we use non ammonical silver staining using two fixatives one hour each, water wash two hours, and then sensitisation with Na2s2o3, developer agno3 - 25 mins, developer na2co3 and finally stop citrate-15 mins total 30 mins time for staining.

Is there a rapid / quick staining protocol giving least background and good sensitivity. NO READYMADE KITS PLEASE.

hoping for a quick reply,

thanks and regards,

exec

hi there
i will tell u the ammmonical silver staining protocal and try it once
1) fixation: 50%methanol,50ul HCHO for 15 min ,change it 3 times
2) wash with dw for 5min 2 times
3) sensitization: 1ml 1M NaOH,49ml dw,mix,add 200mg nano3 mix it u will able to see mud colour add 650ul liqour ammonia shake it well colour get cleard u will able to see like normal water. keep it for 20 min
4) wash with excess dw 10min for 3 times this is the crucial step of forming background.
5) developer: 50ml dw ,1% citric acid 750ul,HCHO 50 ul times depends upon the protein quantity u are using keep it under shaker or shake manually until the develpment of proteins,keep stop solution that is destaining solution over staining develops the background.
all the best

thank you for your response. i will definitely try this method and revert back.

thanks a lot anyways

Hi there,

Here is our Silver Stain Protocol which works very well and is compatable with MS analysis (if that is something you are interested) and is reasonably fast (http://www.pick-n-post.com/files/Alphalyse_Silver_Stain_Protocol.pdf). Hope this helps.

All the best,

AM

exec said:

Can anyone please suggest me a rapid silver staining protocol. we use non ammonical silver staining using two fixatives one hour each, water wash two hours, and then sensitisation with Na2s2o3, developer agno3 - 25 mins, developer na2co3 and finally stop citrate-15 mins total 30 mins time for staining. Is there a rapid / quick staining protocol giving least background and good sensitivity. NO READYMADE KITS PLEASE. hoping for a quick reply, thanks and regards, exec

Did you think using SyproRuby staining.
Much more sensative?
Guy

i would like ot thank everyone for their reply and support. i have started trying the methods and am in the process of standardizing.

thanks a lot for your support.

exec

In any case you would nead any more assistance post us.
Guy