1. Sperm Enzyme Preparation (Group 6) Comparing all Gradient Gel Standards

2. Experiment Objective  Compare all given gradient gel standards and use their relative mobilities to create the best fit mathematical model.  Use best model to compare two gel samples

3. Gathering Data: Gels 1 and 2

4. Hypothesis (Part One)  Standards will be reproducible.

5. Hypothesis(Part Two) Has more proteins than

6. Standards  Measured three times  Calculated relative mobilities  Calculated mean measurements  Best fit mathematical model

7. Mathematical Models  Table of Molecular Weights and the Log of the Molecular Weights: Protein Molecular Weights Log Molecular Weights Myosin200000.005.30 beta-Galactosidase116250.005.07 Phosphorlyase B97400.004.99 Serum albumin66200.004.82 Ovalbumin45000.004.65 Carbonic anhydrase31000.004.49 Trypsin inhibitor21500.004.33 Lysozyme14400.004.15 Aprotinin6500.003.81

8. Best Model Determination RankModels triedStdErrort-invError 1y=a+bX+cX^2+dX^30.07292.57060.1873 2y=a+bX+cX^30.09352.44690.2287 3y=a+bX0.09682.36460.2288 4y=a+bX+cX^2+(lnX)^20.0892.57060.2289 5y=a+bX+cX^20.09742.44690.2383 6y=a+bX+c(lnX)+d(lnX)^20.09572.57060.2461

9. Model with its CI

10. Comparison Chart

11. The Reaction  The expansion of the Vitelline Envelope due to contact with sperm over a 30 minute time period.

12. Comparing Unknowns to Standards  Vitelline Envelope control  Sperm enzyme control  Vitelline Envelope + sperm enzyme (t=0 min)  Vitelline Envelope + enzyme (t=5 min)  Vitelline Envelope + enzyme (t=10 min)  Vitelline Envelope + enzyme (t=15 min)  Vitelline Envelope + enzyme (t=30 min)

13. Results:  Sperm enzyme controls between gels 1 and 2 were similar. SpermEz 5.3248995.232962 5.2261044.893098 4.9387034.844403 4.8800034.805563 4.8052014.746631 4.745294.660919 4.6414044.593366 4.5565224.544095 4.3225214.396276 4.194351 3.937239

14. Results:  Vitelline envelope controls are similar between gels one and two. VE-control 5.0251824.97541 4.8362224.858922 4.6031484.675516 4.5565474.595335 4.5073084.541261 4.3691694.474135 4.373088

15. Results:  In gel 1, the vitelline envelope remains the same throughout the experiment. VE controlt=0t=5mint=10mint=15mint=30min 5.045465.063475.046365.046285.002095.04128 4.852994.868454.864724.857284.806124.84932 4.632424.706204.704174.698044.626574.70021 4.591164.644794.639534.635644.577494.64055 4.548084.609854.599914.592374.524374.56039 4.428734.532894.534814.524914.404744.34733

16. Results:  Vitelline envelope breaks up as the bands become greater. VE controlt=0t=5mint=10mint=15mint=30minEnz Control 4.994575.030095.029135.020495.306365.010155.25677 4.875864.919074.915004.925975.002804.923574.91052 4.697934.843984.844744.851874.906884.845084.86121 4.625464.704644.769034.789144.843934.783174.82238 4.577744.641804.706794.707384.783634.704304.76481 4.519254.566804.613284.655424.701274.637634.68454 4.43210 4.539404.623244.642894.569244.62371 4.556444.602134.456044.58022 4.484254.541593.963954.45204 3.966784.46121 4.27900 3.96145 4.05928

17. Results: Standards

18. Conclusion:  Standards are reproducible.  Sperm enzyme breaks up vitelline envelope. –Number of bands increase. –Small proteins are being detected.