Reagents for flow cytometry

1. PBS 10 X (Stock)

2. PBSA 1 X (Working)

3. Lysing solution 10 X & 1 X

4. PBS-PFA

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1 liter stock of 10x PBS can be prepared by dissolving

• 7.2 g NaCl,

• 1.32 g Na2HPO4 · 2H2O

• 7.52 g KH2PO4

• Make upto 1 L by distilled water.

• pH 7.2 ± 0.2, Store 2-80C in tightly closed bottle

• Dil stock 1 in 10 to a total vol that you would consume in a weak for 1x PBS working

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1 liter stock of 1x PBSA can be prepared by dissolving

• 10g BSA

• 2.5g Sodium Azide

• Make upto 1 L by 1x PBS.

• pH 7.2 ± 0.2, Store 2-80C in tightly closed bottle

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1 litre of Lysing solution stock (10X)

• 89.9g NH4Cl

• 10.0g KHCO3

• 3.7g EDTA Na4

• DW to 1 lit.

• pH 7.2 ± 0.2, Store 2-80C in tightly closed bottle

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100ml Lysing solution working (1X)

• 90 ml DW + 10 ml 10X stock

• Mix well. Store at RT. Discard after 3 days

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PBS – PFA (0.5%)

• PFA 0.5g in 100 ml of 1 X PBS (with azide)

• Dissolve by heating to 65-700C (hot plate with magnetic stirrer is best). Must keep flask/beaker covered tightly with AL foil while heating.

• Filter solution though a 0.2 μ suction filter before use esp for minimal residual disease work.

• Store at 40C for upto 2 weaks