> To: immunology at net.bio.net> From: leew at ph.albany.edu (William Lee)
> Subject: Gey's Solution
> Date: 24 Aug 93 11:28:48 GMT
>> Help! I've lost my Mishell/Shiigi book. If anybody has a recipe
> and protocol for Gey's solution for lysing RBC's, could you
> please email or Fax (518-474-8366) it to me?
>> Also, what are everybody's favorite ways to remove mouse RBC's?
>> Thanks.
>> Bill.
>>The procedure from Mishell/Shiigi is as follows:
Stock A:
NH4Cl 35.0 g
KCl 1.85 g
Na2HPO4-12H20 1.5 g
KH2PO4 0.12 g
Glucose 5.0 g
Phenol red 50 mg
Bring to 1 L
Stock B:
MgCl2-6H20 0.42 g
MgSO4-7H20 0.14 g
CaCl2 0.34 g
Bring to 100 ml
Stock C:
NaHCO3 2.25 g
Bring to 100 ml
1X Gey's solution:
20 parts Stock A
5 parts Stock B
5 parts Stock C
70 parts distilled water
Procedure:
1. resuspend approximatelyt 10e8 pelleted spleen cells in 1 ml of
BSS-5%FCS
2. Add 5 ml of 1x Geys and hold cells on ice for 3-6 min
3. Underlay the cells with 100% FCS and centrifuge at 300 x g for 10
min. Wash once or twice in BSS-5% FCS prior to use.
Hfleit at path.som.sunysb.edu
