First time ELIZA? protocol?
Posted 30 September 2009 - 05:44 PM
I want to do ELIZA for the first time. Can any one help me with the protocol. I have primery abs. No one here can help me with how to do it. Which secondary I should use? Which substrate I need to use to get colour?
can you please kindly help me?
Posted 01 October 2009 - 02:55 AM
Posted 01 October 2009 - 09:27 PM
I am measuring the apoptosis family proteins. I want to check their expression level in cell lysat. I have secondary ab which is HRP conjugate.
Please tell me if you need moreinformation.
Posted 04 October 2009 - 06:39 PM
Thanks for your the website. I was looking for the protocol that you send it. I will try it and ask you Questions accordingly.
Cheers and thanks,
1. From sgt4boston: Tell us the species origin of your primary antibodies (e.g., rat, mouse, rabbit)
2. Do you know the isotype(s) of the primary antibodies you have (e.g., IgG, IgG1, IgG2a, IgG2b, IgM . . . )
I like to use TMB substrate with HRP (e.g., http://www.piercenet...fldID=01041009)
Here is a typical ELISA protocol from abcam (http://www.abcam.com...ource&rid=11422) to get you started with coating/blocking/incubation with antibodies/developing and more . . .
Here is a nice illustration of step by step sandwich ELISA of the type you'll likely be running (http://www.raybiotech.com/elisa.asp)
Hope this is helpful.
Posted 13 October 2009 - 04:46 PM