Gey's Solution

HFLEIT at Path.SOM.sunysb.edu HFLEIT at Path.SOM.sunysb.edu
Tue Aug 24 09:40:16 EST 1993


> 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



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