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

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Presentation transcript:

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

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

Gathering Data: Gels 1 and 2

Hypothesis (Part One)  Standards will be reproducible.

Hypothesis(Part Two) Has more proteins than

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

Mathematical Models  Table of Molecular Weights and the Log of the Molecular Weights: Protein Molecular Weights Log Molecular Weights Myosin beta-Galactosidase Phosphorlyase B Serum albumin Ovalbumin Carbonic anhydrase Trypsin inhibitor Lysozyme Aprotinin

Best Model Determination RankModels triedStdErrort-invError 1y=a+bX+cX^2+dX^ y=a+bX+cX^ y=a+bX y=a+bX+cX^2+(lnX)^ y=a+bX+cX^ y=a+bX+c(lnX)+d(lnX)^

Model with its CI

Comparison Chart

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

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)

Results:  Sperm enzyme controls between gels 1 and 2 were similar. SpermEz

Results:  Vitelline envelope controls are similar between gels one and two. VE-control

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

Results:  Vitelline envelope breaks up as the bands become greater. VE controlt=0t=5mint=10mint=15mint=30minEnz Control

Results: Standards

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