Dear All,
I am working with antibody microarray immunoassy for screening serum proteins. For this assay, I need to dilute the serum samples 1:10 and 1:5. From 1:5 dilution, I measure the protein concentration using BCA essay, which gives variable values for the same serum sample after three consequent measurements. What would be the possible cause for that ? thanks in advance.
Some possibilities:
1. Original sample incompletely mixed.
2. Lack of volumetric reproducibility in liquid dispensing, caused by poor technique or damaged/leaky pipettors.
3. Bubbles in the wells, if using a 96-well plate.
4. Insufficient volume of sample if using a cuvette.
5. Insufficient mixing of solutions.
6. Undissolved solids or turbidity in samples.
7. Poorly functioning spectrophotometer or plate reader.
8. Insufficient incubation time
9. Uneven heating of replicates.
10. Absorbance too close to bottom of standard curve (i.e. too low absorbance), or too far past the linear range (i.e. too high absorbance).
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