Dear Mamta Verma,

Depending on route of administration you can inject different volumes of Ag, specially subcutaneous (tail base) or intradermal you can inject up to 100ul or up to 200 ul in peritoneal cavity. Irrespective of different volume of Ag preparation the emulsification procedure will be same. For more information please follow this article.

Article Immunization of Mice

best wishes

I guess you are not injecting a single mouse. You can prepare all the emulsified antigen for the whole group and even an extra-volume to make it easier to handle. If the volume is too low to pass the two components from one syringe to another until the emulsion is formed, you can also use vigorous vortexing to prepare the emulsion.

As Sunagar Raju write, you can use up to 200ul for ip injection. besides, Prepare antigen in complete Freund's adjuvant using a ratio of 1

volume of antigen to 9 volumes of adjuvant.

Thanks everyone for the suggestions. I finally diluted my protein and used more adjuvant which helped me in making stable emulsion.