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Describe sample 2 (Determine the weight of the monomer, the complex, its polymeric state, and whether or not it contains disulfides). Type your text here: 10% SDS, 100mM DTT, No SDS (Native-PAGE), 10%

          Describe sample 2 (Determine the weight of the monomer, the complex, its polymeric state, and whether or not it contains disulfides). Type your text here: 10% SDS, 100mM DTT, No SDS (Native-PAGE), 10%
        

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Biology for AP Courses
Biology for AP Courses
Julianne Zedalis, John Eggebrecht
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Describe sample 2 (Determine the weight of the monomer, the complex, its polymeric state, and whether or not it contains disulfides). Type your text here: 10% SDS, 100mM DTT, No SDS (Native-PAGE), 10%
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Transcript

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00:02 So describe symbol four of the protein according to three gels and it determined the weight of the monomer, the complex, and its polymeric state, whether or not it contains the sulfide bone.
00:15 So let's take a look at each gel.
00:17 The first one is native.
00:19 Native gel you see two bands.
00:22 So one is about 150 kiloctawton position according to the protein ladder.
00:29 And then the other one is around 50 kilowattelton.
00:32 So that means most likely you'll have a complex which has a total size of 150 plus 50, which is 200 kylodotin protein.
00:47 Now the next step, you'll see a 10 % sds gel.
00:53 So now sds is included to break down certain bond, in our case, non -covalent bonds.
00:59 Now we can see that 150 kylodotin bands disappear.
01:04 Instead, you'll see a thicker band of 75 kallotten.
01:08 And then the other one now drop all the way to 25 -ish.
01:16 And the last one is 10 % sds combined with 100 millimolar of dtt, which is a reducing agent.
01:25 Now, again, 75 klodotan band still exists.
01:31 You still see a band that is 75 kilodont.
01:34 So which means this is probably the monomer.
01:37 Now there's 25 kildalton band now disappear.
01:40 It becomes 12 .5 kilodon.
01:45 So most likely 25 kda divided by 2 equals 12 .5 kda.
01:51 So now let's deduce the polymeric state of the complexes.
02:00 So first of all, what do we know about the sds and dtt? sds is a detergent.
02:07 The very strong detergent breaks all the non -covalent bond.
02:14 The non -covalent interaction includes hydrogen bond, ionic bond, and hydrophobic interaction.
02:32 So all those non -covalent interaction.
02:38 Will be broken down by the inclusion of scs, this strong detergent.
02:44 So if we look at our reaction, you can see 150 kildatenop is gone, so it becomes 75.
02:54 So most likely that the 150 kildolten complex is broken down by the non -covalent bond and c.
03:05 Now it becomes a subunit or a monomer that is 75 kilodontan.
03:13 And for the 50 -kildaden band, this is again a complex.
03:20 Now it's broken down into 25 kilodontent.
03:24 Again, so that means it's broken down into 2 25 kilodotten subunit.
03:31 So then it's going to again be linked by non -covalent bonds.
03:34 So as you can see, we can already tell that there will be at least four subunits out of this protein.
03:43 So 275 kilograms and 225 kilodin, and they're linked by non -cobivalent bonds...
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