# ELISA Testosterone - How to determine hormone level? (Apr/03/2007 )

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Hello,

I am new in ELISA hormone assay. I am doing a research about measuring testosterone level in mice treated by some substance. Unfortunately our ELISA reader cannot create automatic result, thus I need to determine it semi-manually according to the OD. I am already created the standard curve using Excel and now I have no idea how to determine the testosterone level of my samples. I know that it can be harvested from extrapolation based on the standard curve but is there any automatic generation? Because if we generate it manually with ruler the result will be not valid because of the logarithmic scale and the X-axis interval (it is 0; 0.2, 0.5, 1, 2, 6, 16). For instance if the extrapolation is somewhere between 6 and 16, it would be hard to say the hormone level, right? Though there is some minor grid, but still can't be precise as it should be.

-Miss L-

you have to use a polynomial order 2 curve.

-Missele-

QUOTE (Missele @ Apr 3 2007, 04:23 PM)
you have to use a polynomial order 2 curve.

How to do that?

-Miss L-

Ok, I made it but it is not my question.
I want to generate hormone levels of my samples after doing the standard curve.
What should I do?

Thank you

-Miss L-

in excel, right click on your curve , select "add arend curve" or something like that, I don't remember the name in english. it should be the 4th line.

now a new window is opened. in type , choose the righter upper one : "polynomial", or something like that, and in order choose 2.
tell me if it works

-Missele-

in options, did you select show the equation?
then all what you have to do is to calculate the concentration wiht this equation.

-Missele-

Yeah it works well and I got this:

y = -2.288x2 + 3.8584x + 96.862
R2 = 0.9859

Then I have to calculate using this equation, right? Since my data is on Y axis, % (OD/ODo), so I need to figure out the X axis, which is the concentration (ng/mL), right? Whoaa! It sounds nice but how to do that again? I hope it would not be manual opearation..... Shame on me *is getting blushed like over ripe tomato*

-Miss L-

OK, either you find out X, there is a way to do it, but I can't remember,

or, you make a new graph where your concentration is Y

-Missele-

Whaaaa

If I make another graph where concentration is for Y axis, I think it wouldn't change anything, I still have to find put the x things, right?

-Miss L-

QUOTE (Miss L @ Apr 4 2007, 01:58 PM)
Whaaaa

If I make another graph where concentration is for Y axis, I think it wouldn't change anything, I still have to find put the x things, right?

That's right, but this time, the OD/ODo is on the x axis, so you can just plug it into the equation. All your troubles and sad faces will be over

-swanny-

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