Antibody coating protocol

4 posts / 0 new
Last post
Praful
Praful's picture
Antibody coating protocol

Which antibody is used for detection of cry1Ac and cry2Ab insecticidal crystal protein in ELISA ?

what is the protocol for coating of antibody in the ELISA micro plate ?

Where to get these antibodies ?

Braisler
Braisler's picture
I don't know about antibody

I don't know about antibody sources, but coating a plate is pretty simple. You can use Hybond (or similar) plates that are pre-coated to have high protein absorption. Dilute the antibody appropriately in PBS (no BSA or other blocker at this stage) and coat onto plate and cover/seal. I have seen time frames recommended from 1 hour at room temperature to O/N at RT or 4C.

I have also used a 0.1mM solution of sodium bicarbonate to assist in the Ab binding to the plate, but apparently this is not always needed with high absorption plates.

Tirumal Rao
Tirumal Rao's picture
Polynucleotide compositions

Polynucleotide compositions encoding Cry1Ac/Cry1F chimeric O-endotoxins
Disclosed are novel synthetically-modified B. thuringiensis chimeric crystal proteins having improved insecticidal activity against coleopteran, dipteran and lepidopteran insects. Also disclosed are the nucleic acid segments encoding these novel peptides. Methods of making and using these genes and proteins are disclosed as well as methods for the recombinant expression, and transformation of suitable host cells. Transformed host cells and transgenic plants expressing the modified endotoxin are also aspects of the invention.

Tirumal

R Bishop
R Bishop's picture
Praful,

Praful,

I was just at the ASCB meeting and heard about a service called Antibody Dectectives run by AbD SeroTec. They say they will help you find any antibody for free. Im sure they will sell you their's if they have it, but might be worth a try?

Rus