Dear firends
I 've done phage display and I am testing my clones by ELISA. When I was compairing NSB with specific binding ,NSB was too high ! Then I couldn't seperate any positive clones . Someone said to me that my phage titre is too high,is it truth?
What can I do for having better result in ELISA?

i agree that probably the high NSB comes from too high titer.

I have not done ELISA on phage, but I do have done a lot ELISA on other staff. My tips to do good ELISA,

1. the most important thing is to find the proper concentration for both your material and the substrate, to do this, you need to do have serial dilution test

2. for the washing steps, I usually first fill the plate with washing buffer and dump immdiately, then do the washing for a longer time than discribed in protocol.

3. after dumping the liquid in the plate at every step, slam the plate on paper towel real hard to get rid of any trace of liquid

4. when adding anything to the plate, do it as fast as you can, in this assay, good results depends more on the equality than accuracy

5. avoid intercontamination between wells when dumpping the plate