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2 SPF Rabbit, 120 Day Protocol

Procedure
Protocol Day*
Description
Antigen preparation

Peptide synthesis and purification (if required), antigen conjugation
Control serum Collection
Day 0

Primary injection**
Day 1
Primary Immunization with 0.50 mg MAP or 0.25 mg KLH emulsified with Freund's complete adjuvant, SQ 4 sites
1st Booster
Day 14
Boost with 0.50 mg MAP or 0.10 mg KLH emulsified with Freund's incomplete adjuvant.
2nd Booster
Day 28
Boost with 0.50 mg MAP or 0.10 mg KLH emulsified with Freund's incomplete adjuvant.
Serum Collection
Day 42
~25mls per rabbit
3rd Booster
Day 56
Boost with 0.50 mg MAP or 0.10 mg KLH emulsified with Freund's incomplete adjuvant.
Serum Collection, ELISA titer
Day 70
~25mls per rabbit, titer pre, 6, and 10-week bleeds
4th Booster
Day 84
Boost with 1.0 mg MAP or 0.20 mg KLH emulsified with Freund's incomplete adjuvant.
Serum Collection, ELISA titer
Day 98
Large-volume production bleeds (~50mls per rabbit).
ELISA and Shipping
Day 106
Verify disposition of rabbits, Continue*** or Term
Instruction Due Date
Day 120
Termination of animals if instructions not received

* Days listed are approximate (± 2 days).

** Two New Zealand white rabbits will be utilized for each MAP-peptide, conjugated-peptide immunogen, or protein immunogen and are house

in individual cages.

*** Continuation of rabbits is offered on a month to month basis. The standard extension protocol includes one boost, one serum collection, and maintenance for one month. Terminal bleeds may be obtained at any time yielding approximately 50mls of additional sera per rabbit.

For more protocol options, please visit Polyclonal Animal Protocols.

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Procedure	Protocol Day*	Description
Antigen preparation		Peptide synthesis and purification (if required), antigen conjugation
Control serum Collection	Day 0
Primary injection**	Day 1	Primary Immunization with 0.50 mg MAP or 0.25 mg KLH emulsified with Freund's complete adjuvant, SQ 4 sites
1st Booster	Day 14	Boost with 0.50 mg MAP or 0.10 mg KLH emulsified with Freund's incomplete adjuvant.
2nd Booster	Day 28	Boost with 0.50 mg MAP or 0.10 mg KLH emulsified with Freund's incomplete adjuvant.
Serum Collection	Day 42	~25mls per rabbit
3rd Booster	Day 56	Boost with 0.50 mg MAP or 0.10 mg KLH emulsified with Freund's incomplete adjuvant.
Serum Collection, ELISA titer	Day 70	~25mls per rabbit, titer pre, 6, and 10-week bleeds
4th Booster	Day 84	Boost with 1.0 mg MAP or 0.20 mg KLH emulsified with Freund's incomplete adjuvant.
Serum Collection, ELISA titer	Day 98	Large-volume production bleeds (~50mls per rabbit).
ELISA and Shipping	Day 106	Verify disposition of rabbits, Continue*** or Term
Instruction Due Date	Day 120	Termination of animals if instructions not received