• Download an appropriate service request form from our front page:

• Use Polyclonal Antibody Request Form for new polyclonal antibody generation
• Use Monoclonal Antibody Request Form for new monoclonal antibody generation
• Use Miscellaneous Services Request Form for monoclonal antibody reproduction with an existing hybridoma cell line or for other antibody services
• Use Biacore Request Form for Biacore training and instrument usage

1. Fax completed request form to 217-244-0557 (our fax machine does not receive computer fax)

• You will receive confirmation email once we receive your request form
• We will contact you for the progression of your project.
  

Question: What are the antigen requirements?
Answers :

• Quantity: 1.5mg for a new antibody generation with one rabbit or one chicken or 5 mice or 3 rats. If two rabbits or two chickens are used, the amount should be doubled.
• Purity: 90% or better. Contact us if an antigen is less than 90% pure. We might help you with further purification or accept an antigen as it is.
• Concentration: a minimum of 1mg/ml. An accurate determination of an antigen concentration is critical for making a good antibody.
• Buffer: Tris or phosphate buffer with a neutral pH (7-8) are acceptable. No detergents, reducing agents and imidazole should be present in the antigen. Dialyzing antigen in dialysis tubing with 1L buffer stirring overnight at 4C is sufficient for most antigens. The buffer can also be exchanged using Amicon concentrator with several buffer changes. If an antigen does not mixed well with adjuvant due to insufficient dialysis (this happens very often), addition fee will be incurred for further dialysis by the IRC.
• Sterilization: passing through 0.22um filter unit before concentration is determined.
• Shipping: Dry ice or ice pack for overnight service.
• Insoluble antigen is generally acceptable if concentration is well measured. Contact us if you have problem determining concentration with an insoluble antigen.

Answer:
Hybridoma medium can be tested without any dilution. A serum sample needs to be diluted 1:200 to 1:10,000 depending on which bleed and whether the serum is from rabbit or rodent. For rabbit serum, first bleed can be tested at 1:500 to 1:2000 dilutions. Later bleeds to be tested up to 1:10,000. Mouse serum first bleeds can be tested at 1:200 to 1:1000. Later bleeds can be tested up to 1:5000. Chicken serum can be tested following rabbit serum test scheme. Chicken eggs can be treated as chicken serum for dilution.

Immunological Resource Center
Liping Wang, Laboratory Supervisor
264D Burrill Hall, 407 South Goodwin Avenue, Urbana, IL 61801
Phone: (217) 244-0558     FAX: (217) 244-0557    Email: IRC@life.uiuc.edu