protein precipitation

this sight is verry usefull I think if you can view the results for those above questions it is more better
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Go upto 500mM, 750mM and as high as 1M imidazole. it has worked for me with an insoluble HepC protein.

N
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Did you leave it in 250mM imidazole overnight in the freezer...do not do that..dialyse after quickly running fractions on a gel...leaving protein in imidazole will cause protein to precipitate easily..N
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I want to precipitate my proteins that are already solved in Laemmli buffer. Can I use the common methods or is there a special one?

Thanks!
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devi purificare degli enzimi da una soluzione salina attraverso la dialisi,ho bisogno di equazioni riguardanti la dialisi,qualcuno può aiutarmi?
sn una studentessa di ing.gestionale,queste info mi servono xun progetto!
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Dear bio net users

I have extracted dna from wallet nest samples, but I got dna which is contaminated protein. I am looking for a method to precipitate protein contamining my dna. Do you have any suggestion to me? Thank you
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My protein gets precipitate during dialysis within 2 hrs of dialysis. My protein is in phosphate buffer with 300mM NaCl and 250mM Imidazole, I am dialysing it against Trs-Cl buffer with NaCl 100mM.
After dialysis of 2 hrs I observe some precipitate and I observed a huge loss of protein.
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hi all!
i am looking for a protein precipitation method. I did the Acetone precipitation with my protein(animal origin) and got some pellet(enough amount) but when i run it on SDS gel i did'nt get any band.my protein was in 0.3 M EDTA solution, pH-8.can anybody suggest me regarding my problem ? Is EDTA interfere somewhere?
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hi ,
I have expressed one 17 kd protein in bacteria using pqe30 expression vector. i am getting protein as inclusion body.i am trying to purify it under denatured condetion with Ni NTA beads in batch .my problem is that, target protein binds with Ni but it does'nt elute even with 350 mM imidazole. any one can suggest me that what should i do now?
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hello there.

i am precipitating my protein using the ammonium sulfate method. but my enzyme (lipase) activity plunges after dialysis. i tried many attemps but still it gives me the same result. what should i do?

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Hi
Itry to find a method to precipitate proteins from E.coli depends on heating without using organic solvents or ultracentifugation
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Hand-to-Hand Model for Tropomyosin Fast Diffusion.

Semiconductor Mechanism for Diffusion of Tightly-Binding Proteins.

http://actomyosin.narod.ru/h2h-model.htm
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Dear bio net users



I used PET28A plasmid to clone my target gene and

transformed the clone into BL21(DE3).My target protein

expressed as an inclusionbody.I want to raise the

antibody by using this protein.I followed the PET

manual instructions to dissolve and purify the

inclusion body and used the HIS COLUMN to purify the

protein.



My problem is, i am getting my target protein in ELUTE

FRACTIONS ALONG WITH SOME OTHER NON - SPECIFIC

PROTEINS, WHILE WASH BUFFER HAD NO EFFECT ON WASHING

OF NON-SPECIFIC PROTEINS.



Could any one please suggest me any tip to solve this

problem...even i increased the imidazole concentration

to 40mM ( normal range is 20mM of imidazole).

thanking you.

hussain.


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You can try to precipitate your peotein with a salt like ammonium sulphate our with a nonionic polymer, such as PEG. If you chose to work with PEG, you must decide with wiche PEG MW you should work, because it changes with the solutions. You can also try to add somo organic solvents...
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Can you try precipitating your samples with acidified actone (chilled). You can add 10 percent glacial acetic acid to the acetone & try precipitating with this solution such that the final concentration of acetone should be 80 percent in the sample. If you need to remove acetic acid, u can wash the pellet with methanol (60 percent) twice & dry the plle before use. hope it helps you..;
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