SPI-Mark™ Colloidal Gold Reagents
Trouble Shooting (Membranes)
All of the trouble shooting indicators listed above in section 6 apply to
membrane incubations. In addition the following should be considered for
membrane incubations.
(I) Poor labeling
(A) Check transfer system is working.
(B) High background is masking specific staining.
(II) Excessive background
(A) Inadequate blocking of transfer membrane (especially nylon based
membranes). Saturate for longer times at higher temperatures. Use higher
concentrations of BSA in the gold conjugate. Include 0.1% gelatin in the gold
conjugate to reduce binding to sulphur. Wash the membrane very thoroughly
before the incubations.
(B) Spontaneous precipitation of silver due to impure water used for washing
membrane. Use only clean distilled water in clean trays with no contamination.
Use a totally negative control simultaneously.
Inadequate washing of membrane following the gold incubation. Lengthen the
washing procedure in water to remove all salts.
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Thursday May 17, 2012
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