24
edits
Isolation of murine splenocytes: Difference between revisions
From OpenWetWare
Jump to navigationJump to search
Isolation of murine splenocytes (view source)
Revision as of 21:03, 25 September 2006
, 25 September 2006→Materials
No edit summary |
|||
| Line 6: | Line 6: | ||
All materials listed are for use with one mouse. | All materials listed are for use with one mouse. | ||
Supplies | ===Supplies=== | ||
*15ml conical tube | *15ml conical tube | ||
*60mm petri dish | *60mm petri dish | ||
| Line 15: | Line 15: | ||
*50ml conical tube (Protocol B only) | *50ml conical tube (Protocol B only) | ||
Reagents | ===Reagents=== | ||
*DMEM-10 | *DMEM-10 (about 20mL) | ||
**1L DMEM (with 4.5g/L glucose, L-glutamine, sodium pyruvate; from Mediatech, catalog# 10-013-CM) | **1L DMEM (with 4.5g/L glucose, L-glutamine, sodium pyruvate; from Mediatech, catalog# 10-013-CM) | ||
**100mL fetal bovine/calf serum | **100mL fetal bovine/calf serum | ||
**10mL 100x PSG (penicillin G sodium, streptomycin sulfate, L-glutamine; from Gibco, catalog# 10378-016) | **10mL 100x PSG (penicillin G sodium, streptomycin sulfate, L-glutamine; from Gibco, catalog# 10378-016) | ||
***sterilize using 0.2μm filter; store at 4° | ***sterilize using 0.2μm filter; store at 4° | ||
*ACK lysis buffer | *ACK lysis buffer (1mL) | ||
**1L deionized H2O | **1L deionized H2O | ||
**8.29g NH4Cl | **8.29g NH4Cl | ||
| Line 28: | Line 28: | ||
***pH solution to 7.2-7.4; sterilize using 0.2μm filter; store at 4° | ***pH solution to 7.2-7.4; sterilize using 0.2μm filter; store at 4° | ||
*70% ethanol | *70% ethanol | ||
*trypan blue solution | *trypan blue solution | ||
Equipment | ===Equipment=== | ||
*scissors | *scissors | ||
*forceps | *forceps | ||
