Well, your not alone! ( :
I was going to post the steps before but i thought it would be to much. But, I'm post them so you can have a look see...Line 16 through 18 get to the heart of what i need to find. Trying to find what volume to multiple by is a bit tricky too. Mixed and diluted the enzymes with buffers and what not.
Extraction and Quantification of Acid Phosphatase Enzyme from Wheat
Germ
1. Obtain a 15 mL conical centrifugation tube and add 0.5 grams of raw
wheat germ to the tube.
2. Add 5 mL of 0.05M Sodium Acetate buffer, pH 4.5. Cap and shake the
tube to disperse the wheat germ into the buffer
3. Using the sonicator, disrupt the suspended wheat germ by sonicating for
30 seconds.
4. Shake the tube again and with a plastic transfer pipet, remove about 1 mL
of the suspension into a new microcentrifuge tube.
5. Centrifuge on high for 30 seconds and place tube in a tube rack. You will
be using the supernatant (which should contain acid phosphatase) in the
next experiment.
6. We need two rows of 6 wells on your plate. Mark one as pure enzyme and
the other as extract
7. Into both rows add 50 uL of 1M NaOH to each well.
8. Obtain two clean microcentrifuge tubes, label one pure enzyme and the
other as extract.
9. Into each tube, add 600 uL of 0.05M Sodium Acetate buffer, and 100 uL
of the substrate solution.
10. Notify your timer to pay attention!
11. Quickly but accurately add 60 uL of pure enzyme to the pure enzyme tube
and 60 uL of the extracted enzyme to the extracted enzyme tube.
12. Immediately mix well by shaking, start the timer counting up,
immediately remove 100 uL of each tube and put it in the first wells of
each row, combining it with the NaOH already in the well. This is a
time=0 time point
13. Repeat taking out 100uL samples from each tube and putting it in the
appropriate wells at time=2minutes, =4 minutes, =6 minutes, =8 minutes,
and finally, time=10 minutes.
14. On the plate reader, record absorbance values for each well at 405 nm.
15. plot and X-Y plot using the absorbance (on the Y) vs. time (on the X) and
do linear regression through the points.
16. Analyze the linear regression data to obtain the slope value (m) for each
line. This values tells how fast the reaction was proceeding. The more
enzyme, the faster the rate. We can use these values to quantify how
much enzyme is present in the reaction tubes.
17. We added 50 uL of pure active enzyme to the tube to produce the rate
observed in the pure enzyme timed reaction. The concentration was 1
mg/ml. How much active enzyme was in this tube?
18. By setting up a ratio between the slope/enzyme ug to the slope of the
extract reaction, we can determine how much active enzyme was in the
extract.
)
obviously I'm misunderstanding something here 
)