Biology-Online • View topic - Help with the design of an ELISA experiment
Login

Join for Free!
121775 members
Advertisement
Advertisement
Advertisement


Help with the design of an ELISA experiment

Discussion of all aspects of biological molecules, biochemical processes and laboratory procedures in the field.

Moderator: BioTeam

Help with the design of an ELISA experiment

Postby bemardus » Tue Apr 27, 2010 10:39 pm

Hi guys,

So I need help with an ELISA experiment. I need to determine the concentration of IgG in serum and what percentage of the cellular protein in said serum is a result of IgG. What I have to work with is mouse serum, mouse serum albumin, mouse IgG, rabbit anti-mouse IgG, goat anti-mouse IgG (biotin and FC-HRP), goat anti rabbit IgG (biotin and ap-conjugated), goat anti-mouse IgG HRP (gamma chain), rabbit anti mouse serum. I haven't an idea where this should begin. Plus what all these antibodies are used for. Any help would be awesome. Thanks.
bemardus
Garter
Garter
 
Posts: 3
Joined: Tue Apr 27, 2010 10:26 pm

Postby Jillo725 » Tue Apr 27, 2010 11:27 pm

You'll be doing a sandwich elisa test. wikipedia it. it's got a great description and picture to go along with it. all proteins are accounted for in this test.
"There is no adequate defense, except stupidity, against the impact of a new idea."
— Percy Williams Bridgman, US physicist
Jillo725
Garter
Garter
 
Posts: 31
Joined: Tue Apr 27, 2010 5:17 pm
Location: Evans, GA

Postby bemardus » Wed Apr 28, 2010 4:18 am

Thanks Jillo725.

So guys am curious now though what all these antibodies are used for. After looking at wikipedia I have an idea of what to do but not what to use for each step. So I'm guessing I'd used mouse IgG as the capture antibody. Then rabbit anti-mouse IgG as the would be antigen. Following that I'd use goat anti-rabbit IgG to attach to the rabbit anti-mouse IgG. And I'd use goat anti-mouse IgG FC-HRP. Is this at all right?

With that what's the difference between ap-conjugate and biotin? And FC-HRP and gamma chain HRP? Which of these do I choose? And what's the purpose of mouse serum albumin and rabbit anti-mouse serum?

Sorry for all these questions. I'm totally lost here though. Thanks for the help though.
bemardus
Garter
Garter
 
Posts: 3
Joined: Tue Apr 27, 2010 10:26 pm


Re: Help with the design of an ELISA experiment

Postby Jillo725 » Wed Apr 28, 2010 1:50 pm

Not sure about everything. just look for the definitions of those. it could be that the teacher listed extra proteins that are not used. I know that FC is the stem region of the antibody. I think (if i remember right) that the HRP is some kind of human protein....or fluorescence i can't remember, we're going back about a year and a half here =p
"There is no adequate defense, except stupidity, against the impact of a new idea."
— Percy Williams Bridgman, US physicist
Jillo725
Garter
Garter
 
Posts: 31
Joined: Tue Apr 27, 2010 5:17 pm
Location: Evans, GA

Re: Help with the design of an ELISA experiment

Postby savvygirl » Fri May 07, 2010 2:13 am

hehe

Is this someone in Dr. Bob's Class?

:)
savvygirl
Garter
Garter
 
Posts: 1
Joined: Fri May 07, 2010 2:09 am


Return to Molecular Biology

Who is online

Users browsing this forum: No registered users and 0 guests

Advertisement