Truncated Protein on SDS-PAGE_________

sarim - 08/22/11 21:34

#608763

http://www.iovs.org/content/41/9/2466/F1.large.jpg just wanted to share this. e.g. due to the "Non -sense Mutation(STOP Codon)", note that "Truncated Protein" Band appear at the Bottom on SDS-PAGE due to it's smaller size. * Re:Truncated Protein on SDS-PAGE_________
technodoctor - 08/22/11 21:43

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#2455460
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thankx :) * Re:Truncated Protein on SDS-PAGE_________

sharif0812 - 08/22/11 21:45

#2455463
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hey thanks sarim for this---do u kno why the truncated one is on the bottom?? * Re:Truncated Protein on SDS-PAGE_________
technodoctor - 08/22/11 21:58

#2455470

@sharif0812....the truncated one is smaller in size and therefore would travel farther. The normal protein is heavier and therefore remains closer to original position. I hope you know that these proteins travel towards the opposite charge in electrical feild...they being -vely charged travel towards + electrode... the lighter the protein, the farther it will go * Re:Truncated Protein on SDS-PAGE_________
step1saga - 08/23/11 02:51

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#2455591

Thanks sarim!! I think you should get be the only person to get a 300 ...and I m sure you'll come pretty darn close :) (also saying this based on your nbme explanations--thanks again) * Re:Truncated Protein on SDS-PAGE_________
step1saga - 08/23/11 03:00

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#2455593

300/99* Im gonna add something about sickle cell on sds page HbA = Is valine( which is a hydrophobic AA)-- so it goes towards the positive charge in the electrodee HbS = neutral HbC= is lysine (positively charged AA)- moves towards the negative electrode * Re:Truncated Protein on SDS-PAGE_________
step1saga - 08/23/11 03:03

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#2455595
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does SDS page involve charged electrodes? or just argose gel? * Re:Truncated Protein on SDS-PAGE_________
sarim - 08/23/11 04:03

#2455598

@step1saga thanks for ur nice words.wish u all the best.

SDS-PAGE: -is an Electrophoresis as the name states(SDS-polyacrylamide gel Electrophoresis), so it does involve + and - poles.......BUT separate the protein strands by their "SIZE" (and not by their Intrinsic Charge)

-the gel is prepared with wide range of average Pore sizes -u can read about SDS-PAGE on net, it will help u solve Qs that otherwise seem weird. -Here are some Diagrams in the following links ......if u would like further discussion, i'll be glad to join. http://www.molecularstation.com/images/protein-gel-electrophoresis.jpg http://ww2.chemistry.gatech.edu/~lw26/bCourse_Information/4581/techniques/gel...ect/gel.jpg http://talon.niagara.edu/~391s08/giacomini/images/sds_page.png http://biochem.co/wp-content/uploads/2008/08/sds-page-gel-and-graph.png Note that the standard molecular wt (on the Lt side) are given as reference and they are spaced out from Top(larger fragments) towards Bottom(Smaller Fragments) http://www.personal.psu.edu/jms5704/blogs/simmons/1476-7961-4-12-1.jpg good luck.

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odin - 08/23/11 19:49

#2456028
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Thanks, sarim! Very helpful. * Re:Truncated Protein on SDS-PAGE_________

step1saga - 08/23/11 20:52

#2456050
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thanks for the links sarim :) * Re:Truncated Protein on SDS-PAGE_________

sarim - 08/23/11 21:09

#2456051

:) thanks Note how SDS give out Negative charge to all protein molecules (Masking the protein Intrinsic charge) http://www.ebioe.com/uploadfile/2010/0409/20100409022030595.jpg http://www.gibthai.com/userfiles/image/technote/Western%20Blot/Western...-Blot-2.jpg

step1saga - 08/23/11 21:19

*they can also put it sideways http://www2.montana.edu/cftr/images/sds_page5.gif yeah that makes charge to be completely inconsiderable or negligible to have any significant effect so they would definitely not show sickle cell anemia on a SDS. Since defiinitely determined by size. I almost turned my laptop 90 degrees to see the one sideways..
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psychmledr - 08/23/11 22:57

#2456109

Thank you so much :)