Reagents
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Sodium Borohydride (NaBH4) - 1 mg/ml
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Carbonate:Bicarbonate Buffer, pH 9.6
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Sodium Periodate (NaIO4) - 1 mM, 2.14 mg/10 ml Acetate Buffer
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10mM Acetate Buffer, pH 4.5
0.82 g sodium acetate in 90 ml diH2O.
Adjust to pH 4.5 with glacial acetic acid. Q.S. to 100 ml with diH2O.
Procedure
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To one-half the plate add 100 ml/well periodate. To the other half add 100 ml/well acetate buffer.
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Incubate 2 hr at 37oC.
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Wash 2X with 0.1M bicarbonate buffer.
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Add 100 ml/well sodium borohydride.
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Incubate 30 min, RT
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Wash plate 2X with PBS
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Block plate as required by assay protocol.
Notes
In fixed cell ELISAs. do not use carbonate wash before substrate