Sperm Enzyme Preparation (Group 6)
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Transcript of Sperm Enzyme Preparation (Group 6)
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Sperm Enzyme Preparation (Group 6)
Comparing all Gradient Gel Standards
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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
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Gathering Data:
Gels 1 and 2
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Hypothesis (Part One)
Standards will be reproducible.
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Hypothesis (Part Two)
Has more proteins than
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Standards
Measured three timesCalculated relative mobilitiesCalculated mean measurementsBest fit mathematical model
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Mathematical Models
Table of Molecular Weights and the Log
of the Molecular Weights:
Protein
Molecular Weight
sLog Molecular
Weights
Myosin 200000.00 5.30
beta-Galactosida
se 116250.00 5.07
Phosphorlyase B 97400.00 4.99
Serum albumin 66200.00 4.82
Ovalbumin 45000.00 4.65
Carbonic anhydrase 31000.00 4.49
Trypsin inhibitor 21500.00 4.33
Lysozyme 14400.00 4.15
Aprotinin 6500.00 3.81
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Best Model Determination
Rank Models tried StdError t-inv Error
1 y=a+bX+cX^2+dX^3 0.0729 2.5706 0.1873
2 y=a+bX+cX^3 0.0935 2.4469 0.2287
3 y=a+bX 0.0968 2.3646 0.2288
4 y=a+bX+cX^2+(lnX)^2 0.089 2.5706 0.2289
5 y=a+bX+cX^2 0.0974 2.4469 0.2383
6 y=a+bX+c(lnX)+d(lnX)^2 0.0957 2.5706 0.2461
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Model with its CI
0.1 0.2 0.3 0.4 0.5 0.6 0.7 0.8 0.9
4.0
4.5
5.0
5.5
RmMean
logM
wlogMw = 5.96754 - 5.56606 RmMean
+ 9.56238 RmMean**2 - 6.92265 RmMean**3
S = 0.0729144 R-Sq = 98.5 % R-Sq(adj) = 97.6 %
Regression
95% CI
Regression Plot
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Comparison Chart
3.5000
3.7000
3.9000
4.1000
4.3000
4.5000
4.7000
4.9000
5.1000
5.3000
5.5000
0 0.2 0.4 0.6 0.8 1
Rm
Lo
gM
w PreVs.RmMean
StdLogMwVs.RmMean
StdRmGel1
StdRmGel2
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The Reaction
The expansion of the Vitelline Envelope due to contact with sperm over a 30 minute time period.
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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)
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Results:
Sperm enzyme controls between gels 1 and 2 were similar.
SpermEz SpermEz
5.324899 5.232962
5.226104 4.893098
4.938703 4.844403
4.880003 4.805563
4.805201 4.746631
4.74529 4.660919
4.641404 4.593366
4.556522 4.544095
4.322521 4.396276
4.194351
3.937239
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Results:
Vitelline envelope controls are similar between gels one and two.
VE-control VE-control
5.025182 4.97541
4.836222 4.858922
4.603148 4.675516
4.556547 4.595335
4.507308 4.541261
4.369169 4.474135
4.373088
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Results: In gel 1, the vitelline envelope remains the same
throughout the experiment.
VE control t=0 t=5min t=10min t=15min t=30min
5.04546 5.06347 5.04636 5.04628 5.00209 5.04128
4.85299 4.86845 4.86472 4.85728 4.80612 4.84932
4.63242 4.70620 4.70417 4.69804 4.62657 4.70021
4.59116 4.64479 4.63953 4.63564 4.57749 4.64055
4.54808 4.60985 4.59991 4.59237 4.52437 4.56039
4.42873 4.53289 4.53481 4.52491 4.40474 4.34733
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Results: Vitelline envelope breaks up as the bands
become greater.VE control t=0 t=5min t=10min t=15min t=30min Enz Control
4.99457 5.03009 5.02913 5.02049 5.30636 5.01015 5.25677
4.87586 4.91907 4.91500 4.92597 5.00280 4.92357 4.91052
4.69793 4.84398 4.84474 4.85187 4.90688 4.84508 4.86121
4.62546 4.70464 4.76903 4.78914 4.84393 4.78317 4.82238
4.57774 4.64180 4.70679 4.70738 4.78363 4.70430 4.76481
4.51925 4.56680 4.61328 4.65542 4.70127 4.63763 4.68454
4.43210 4.53940 4.62324 4.64289 4.56924 4.62371
4.55644 4.60213 4.45604 4.58022
4.48425 4.54159 3.96395 4.45204
3.96678 4.46121 4.27900
3.96145 4.05928
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Results: StandardsLogMw Vs. Rm
3.5
3.7
3.9
4.1
4.3
4.5
4.7
4.9
5.1
5.3
5.5
0.05 0.15 0.25 0.35 0.45 0.55 0.65 0.75 0.85 0.95
Rm
Lo
gM
w
Rm1
Rm2
Rm3
Rm4
Rm5
Rm6
Rm7
Rm8
Rm9
Rm10
Rm11
Rm12
Rm13
Rm14
Rm15
Rm16
Rm17
Rm18
Rm19
Rm20
Rm21
Rm22
mean
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Conclusion:
Standards are reproducible.Sperm enzyme breaks up vitelline envelope.
– Number of bands increase.– Small proteins are being detected.