Technical considerations – Bio-Rad Matrix-Prepared Surfaces User Manual

Page 4

Advertising
background image

3

© 2007 Bio-Rad Laboratories, Inc.

Technical Considerations

n

If your sample has salt or urea contamination, proceed directly

to Protocol 2. Otherwise, perform Protocol 1 first. If there is
insufficient signal after reading the arrays, a cleanup step is
recommended: Add 5 µl of 0.1% trifluoroacetic acid (TFA) to
the spot; do not agitate; remove the 0.1% TFA droplet after
30 sec; and add 5 µl 25% acetonitrile (ACN) and 0.1% TFA to
the spot. Allow to dry, and read the arrays again

n

With ProteinChip SEND ID arrays, it is essential that you mix

your sample with a solution of 50% ACN

and

0.2% TFA at a

1:1 (v/v) ratio before adding to the spot

n

Ideally, the final concentration of ACN after dilution is 25%;

a final concentration of greater than 40% ACN is not compatible
with the ProteinChip SEND ID array

Factors That Can Result in Weak Signal

n

We do not recommend washing on-spot (by pipetting the

sample up and down) because this can reduce signal

n

Weak signal can be improved by adding ACN to your sample

as described in Protocol 1

n

A predominant peak at mass-to-charge ratio (m/z) 211

indicates sodium contamination, and you should perform
sample cleanup as described in Protocol 2, step 5. This
contamination can suppress sample peaks and result in
weak signal

Advertising