After seeing how little of the sample is consumed to acquire the desired number of beads, I thought we may be able to stretch the kit by lowering the number of beads per sample. Hopefully this wouldn't augment the results. I was wondering if anyone had stretched their Miliplex kit.
We purchase our beads from Bio-Rad and use half the suggested amounts. It produces great results!
We purchase our beads from Bio-Rad and use half the suggested amounts. It produces great results!
I agree, you can lower the amount of beads (antibody concentration), but make sure you keep the volume the same as not to introduce a calculation error.
Gregory S Whitehead and Colin Reardon, I have Question for you.
How do you do it exactly?
Do you cut the volume in half for all the steps?
Or You just reduce the concentration by one-half of beads and keep volume same? And if you do this.. Do you reduce the concentration of Ab and S-PE too?
It would be really nice if you provide some detail about it.
Yes, as Eva Morshi answered, we lower the amount of beads, antibody concentration, in half for both the coupled beads and detection Ag, but we add the same volume (50 or 25 ul) to the 50 ul sample. You can or cannot reduce the S-PE - works either way!
HI Gregory S Whitehead, Thank you very much for your reply.
Do you decrease concentration of the analysts in the standard curve (S1 to S8) to half or you keep same?
Question arises in my mind is that if I don't decreases the concentration of the analysts in standard curve the plateau will be reached before? There will be shift for other points too.
I will be pleased to have your comment.
yes, decrease the concentration for the standards as well: Everything is treated the same!
Hey Gregory thanks for your answer...I have one more particular question. It will be good, if you can help. Concentration of analyst entered for the standard in the software should be half (for S1, S2...so on) OR keep it same because the ratio is same.
Sorry for my last answer - the concentration of standards will remain the same, but just add lower amount of coupled beads and detection Ag to them; thus, keeping the concentration of analyst the same.
So far I have been preapring beads at a density of 200beads/ul and then adding 25ul per well to get 5000 beads (200beads/ul*25ul) per well. I have not tried it yet but I feel this bead number can be very well lowered to as low as 2500 beads per well, i.e., to use either 50beads/ul and then add 50ul per well or use 100beads/ul and then add 25ul per well.
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