ELISA methods - needing information/papers (Dec/02/2005 )
Hi Guys,
I've been reading lots here about ELISAS which has been really helpful, however I've noticed that there seem to be two methods currently being queried.
The first is the Sandwich ELISA technique (Ab, block, protein, ab, measure), but I've seen otjher people enquiring about another technique where you coat the well with the protein, then add a primary and secondary ab. I need to know if this is a valid technique (i.e one which has featured in a paper) and if there is a protocal availible for it. I am planning to filter my total protein to get a very small range of molecular weights and am wondering if this might be a suitable alternative to the Sandwich ELISA technique.
Any advice is greatfully recieved, especially links to papers featuring this.
Thanks,
Rosie
I've been reading lots here about ELISAS which has been really helpful, however I've noticed that there seem to be two methods currently being queried.
The first is the Sandwich ELISA technique (Ab, block, protein, ab, measure), but I've seen otjher people enquiring about another technique where you coat the well with the protein, then add a primary and secondary ab. I need to know if this is a valid technique (i.e one which has featured in a paper) and if there is a protocal availible for it. I am planning to filter my total protein to get a very small range of molecular weights and am wondering if this might be a suitable alternative to the Sandwich ELISA technique.
Any advice is greatfully recieved, especially links to papers featuring this.
Thanks,
Rosie
I think you are refering to the Direct ELISA. This is not as accurate as Sandwich ELISA.
Pierce and Biosource both have some great ELISA info on their websites, with references listed...I would suspect any vendor that sells alot of ELISA stuff probably also has some good info and tips for you
I've been reading lots here about ELISAS which has been really helpful, however I've noticed that there seem to be two methods currently being queried.
The first is the Sandwich ELISA technique (Ab, block, protein, ab, measure), but I've seen otjher people enquiring about another technique where you coat the well with the protein, then add a primary and secondary ab. I need to know if this is a valid technique (i.e one which has featured in a paper) and if there is a protocal availible for it. I am planning to filter my total protein to get a very small range of molecular weights and am wondering if this might be a suitable alternative to the Sandwich ELISA technique.
Any advice is greatfully recieved, especially links to papers featuring this.
Thanks,
Rosie
I think direct ELISA is more accurate,good luck
hi
i have been using the method u mentioned.
I have coated wells with Antigen overnite.
Then blocked it..serum(Ab)...Biotinylated Ag...Streptavidin HRP...TMB..
it is found to be effective..
Check it out
vvi