Third+Mouse+CTL+Experiment

6 Mice arrived on 4/24/12,( Balb/C) which had been put in 40A in Bio B. These Balb/c mice cannot be used for Ova immunization! I must use C57BL/6J mice for the immunization. I sent an e-mail to Kathy to order these C57BL/6J mice. https://mail.google.com/mail/u/0/?ui=2&shva=1#sent/137b9414ea0d4d44

The cells I will use with the splenocytes from these mice are B16F10 cells: B16F10 Cells started 7-22-12

C57BL slash 6J mice arrived 6-12-12 There are 5 mice. 4 will be immunized, and 1 will be a control.

Identity of Ova protein for protein immunization: Ova Protein Concentrated 10K HW 1-28-11 in 1XPBS at Ova protein box -20 C Identity of Ova plasmid for genetic immunization: pCMVi LS Ova 9/21/11 337.2 (ug/mL?) at CTL 6-13-11 -20 C

Tentative schedule: Genetic immunization day 1 with pCMVi-LS Ova 2 bullets per mouse (4 mice); 1 ug pCMVi-LS Ova plasmid and 1 ug CpG C per bullet Genetic immunization day 3 with pCMVi-LS Ova 2 bullets per mouse (4 mice); 1 ug pCMVi-LS Ova plasmid and 1 ug CpG C per bullet Protein boost day 17 with 200 uL of 0.05 mg/mL SAS adjuvant 0.025 mg/mL Ova protein (see 3-30-12 of notebook for last protein boost)

Actual schedule Genetic immunization (6/18/12) day 1 with pCMVi-LS Ova 2 bullets per mouse (4 mice); 1 ug pCMVi-LS Ova plasmid and 1 ug CpG C per bullet. 1 mouse was kept as a naive mouse, and it's ear was hole punched. Genetic immunization (6/20/12) day 3 with pCMVi-LS Ova 2 bullets per mouse (4 mice); 1 ug pCMVi-LS Ova plasmid and 1 ug CpG C per bullet Protein boost (morning of 7-3-12) day 16 with 200 uL of 0.05 mg/mL SAS adjuvant 0.025 mg/mL Ova protein <- (I think this must be 0.5 mg/mL SAS 10-3-12) Cheek Bleed of Mice on 7-12-12 Protein boost (afternoon of 10-2-12) of 1 mouse (marked with a black marker) with 200 uL of 0.5 mg/mL SAS adjuvant 0.05 mg/mL Ova protein Protein boost (afternoon of 10-15-12) of 1 mouse (marked with a black marker) with 200 uL of 0.5 mg/mL SAS adjuvant 0.05 mg/mL Ova protein Splenectomy of one mouse 10-20-12 (only the naive mouse is left)
 * Protein was 0.1 mg/mL protein prepared from "Ova Protein Concentrated 10K HW 1-28-11 in 1XPBS 6.47 mg/mL at Ova Protein box -20 C". Note that one of the mice partially leaked when being immunized. We injected a little more solution into this mouse, and then I had to come back upstairs from the vivarium to prepare a little bit more protein.