Primary antibody and Sodium azide
Posted 10 August 2009 - 09:49 PM
Days a few back I faced problems in getting signals from my blot, while I diluted the primary ab in 3% BSA and with 0.1% NaN3. While diluting the same ab with similar condition, only reducing the NaN3 to 0.01% resulted the clear signal in the band.
Does any one know if NaN3 has a deleterious effect on the primary an other than being an anti-bacterial preservative?
Posted 11 August 2009 - 04:02 AM
Posted 11 August 2009 - 07:04 AM
the dedection is based on HRP. I do three washes with TBS-T, 5 min each before adding HRP-conjugated secondary ab. Probably the washes are not enough. I'll check increasing the wash.