How To Interpolate A Standard Curve In GraphPad Prism

In this video tutorial, I will show you how to interpolate a standard curve by using GraphPad Prism.
In the example, I have performed an ELISA experiment where I need to calculate the concentrations of my samples with known absorbance values, through the use of a standard curve. The standard curve in this example is that of a hyperbola relationship.
GraphPad Prism version used: 6
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[@StevenBradburn]

THE ONLINE GUIDE
toptipbio.com/standard-curve-graphpad/

[@mlgc95]

This saved my life and my Masters's thesis work. Thank you so much!

[@claudiacarrillo6315]

thank you sm, you just saved my life and career

[@fishflavoredicecream]

Hi, may I ask, how does this take into account the dilution of serum samples? For example, what if on a single plate with two treatment groups and a standard, one treatment group sera were diluted as 1:500 and the other as 1:200? Can they still use the same standard?

[@sanjuktachakraborty8814]

Thank you for the tutorial. Helped me a lot.

[@StevenBradburn]

Very welcome

[@mikenaughton]

What about the curve performance? How do you find out or display the concentration of each calibration point? If you were running duplicates I'd want to see the %RE (accuracy) to the spiked conc and the %CV (precision) of replicates for each level. Then the acceptability of the analytical run can be determined (assuming 20 - 30% acceptance criteria).

[@peacefulcali]

Thank you for the video. Can you get LLOQ and ULOQ data from this as well?

[@benysmart1643]

thanks for all tips

[@Albatross039]

very clear explanation, thank you

[@gislenegil9585]

Thank you. It helped me a lot.

[@meteorcube]

Thanks!

[@WanapornIttiprasertTanno]

Thank you!

[@StevenBradburn]

You're welcome!

[@prajithnambiar4335]

hi how to interpolate in graphpad prism 5

[@StevenBradburn]

Hi Prajith, I believe this feature is only available in Prism 6 onwards

[@dr.asaadaladlaan1079]

IT IS NOT ACCURATE, TRY PUT THE SAME ABSORBANCE OF THE S1 OR S2 AS UNKNOWN. YOU WILL SEE DIFFERENT CONCENTRATION. LET SAY THAT S1 (30 NG/ML) WITH OD (0.667) WHEN YOU PUT THE SAME OD 0.667 AS AN UNKNOWN THAT WILL GIVE YOU CONCENTRATION 32.45 AND NOT 30. IT SHOULD BE 30 NG/ML SINCE THE OD IS THE SAME 0.667. THAT RESULT MIGHT SHIFT RESULTS TO POSITIVE INSTEAD OF NEGATIVE.

[@juliamadeleineheinze2932]

The curve you receive is an approximation, that is calculated to fit all points as best as possible. In an ideal world, your curve would go through each measured dot precisely. But in reality, not every dot fits perfectly onto the standard curve, which is why you receive slight differences when plugging the concentration of your standards in. The more preceise you work (hands on), the more precise your standard curve will be and the better you will appoximate the concentration of your samples.