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Formalin administration may lead to an increase of NO which in turn would lead to formation of cyclic guanosine monophosphate (cGMP) by activation of guanylyl cyclase

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A80-229AP, Bethyl Laboratories, Inc.), diluted in PBS/T/M (pH 5.5 or pH 7.4), was added for 1 h at room temperature, followed by a washing step and detection with 1 mg/mL (100 L/well) of p -Nitrophenyl Phosphate Substrate (Sigma-Aldrich) dissolved in diethanolamine buffer (pH 9.8) (Sigma-Aldrich)