Standard Operating Procedure

 

Neuraminidase Treatment

 

Hazards:

NEURAMINIDASE: harmful if inhaled or absorbed through the skin and may be harmful if swallowed. Will cause irritation to the mucous membranes and upper respiratory tract and may cause allergic respiratory and skin reactions. Repeated exposure may cause asthma.

Protection:

An alert and conscious mind.

Waste:

drain

Spill Clean-Up:

 

Procedure:

• Use 1-2x10⁶ cells

 

1.     Wash with RPMI.

2.     Warm cells in 37^oC water bath for 10-15min.

3.     Add neuraminidase (from Vibrio cholera; Sigma N6514) 0.016units/2x10⁶ cells.

4.     Incubate at 37^oC for 20min.

5.     Quench reaction with 10x volume of RP10

6.     Wash 3x with RP10.

7.     Stain an aliquot of untreated (i.e. RPMI alone) and neu treated cells with PNA-FITC (1:1000) to check for effectiveness of the neu treatment.