70 DAY ANTIBODY PRODUCTION PACKAGE - Rats and Mice
This protocol replaces the Traditional Antibody Production Protocol that we offered. We have kept the the immunization and bleeding schedule the same. Please keep in mind that antigen in gel can not be injected intraperitoneal. If your antigen is in a gel, we can still follow this protocol, but all injections will be subcutaneous or you can select the 91 day protocol.
Day 0 -
Preimmune Bleed, about 0.1ml of sera. Preimmune bleed is stored until the first immune bleed is sent.
Inject 50 to 100ug intraperitoneal in Complete Freund's Adjuvant (FCA). Antigen supply should be 1mg/ml or higher.
Day 14 -
Inject 25 to 50ug intraperitoneal in Incomplete Freund's Adjuvant (IFA)
Day 28 -Inject 25 to 50ug intraperitoneal with IFA.
Day 42 -
Bleed. About 0.1ml sera for mice and 0.5ml for rats - This bleed will be sent to you along with the preimmune bleed.
Day 56 -
Inject 10 to 25ug intraperitoneal with IFA
Day 70 - Select one of the three options below:
For CLOSED projects - Bleed at day 70 - Rat or Mouse is then euthanized / discard (lowest cost). The bleed is then sent to you.
For CLOSED projects - The Rat or Mouse is exsanguinated (final bleed) at day 70. The exsanguination sera is then sent to you.
For OPEN projects - Bleed - This bleed is then sent to you.
After Day 70 for OPEN projects -
You may select one of the project extensions below. If you do not specify a project extension, we will automatically begin Extension A. Keep in mind that all procedures and maintenance after day 70 will be charged according to the Custom Polyclonal Antibody Production Price Schedule.
We will boost every four weeks with 10-25ug of antigen intraperitoneal with IFA. We will do a large bleed two weeks after each boost and ship. We will continue to boost and bleed every four weeks until you contact us.
Extension B:When the antisera contains sufficient antibody titer and large volumes of sera are needed, we can start production bleeds. We do production bleeds every seven days. We will continue with production bleeds until you contact us or the health of the animal requires us to stop production bleeds. The volume of serum collected for large or production bleeds depends on the weight and the general health of the rat or mouse. Generally, bleed volumes will not vary from the bleeds previously drawn. During this time, we boost the animal every four weeks with 10-25ug of antigen intraperitoneal with IFA. You may also request that we not do any boosts after the initial immunization scheme outlined above since some antigens and some animals may respond negatively to boosts during the period of peak antibody activity. Boosts may be required only when titer drops. It is up to the investigator to decide and let us know if we should discontinue boosts as our standard schedule provides for boosts every 28 days. Shipping for production bleeds is every four weeks unless you request otherwise. Please keep in mind, unlike our other species, rats and mice do not tolerate many cycles of production bleeds.
Extension C:Some investigators prefer to "wait and see." after day 91. Extension C is hold without boosting and bleeding. We will hold until you tell us to do something else.
When you have collected enough antisera, you will need to contact us to terminate the project. You may request that we terminate by:
When requesting termination, you must confirm the species and animal number. Written confirmation by email may be requested for terminal procedures.
Spleen Harvest:At the time of exsanguination, we are able to harvest the spleens. BALB/c mice and Long Evans rats are used for antibody production. Please specify the conditions of which you want the spleen. Additional charges apply for spleen harvests.
You may design your own project extension. Let us know what we can do for you!