Third Mouse CTL Experiment

2015-01-13

azim58 - 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&amp;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)

* 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.
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)

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