Question

which methods would i use to purify the protein 10 from agbooth?

Quit Separation PAGE Fractions Help pH 80K 60K 50K- 40K- 30K- 20K- 5K 10K- 5K 2D-PAGE of initial mixture 4 Record pH Ammoniurm sulfate frectionation Heat treatment Gel filtration ton exchange chromatography 80K- 60K 50K 40K- 30K- 1-Dimensional PAGE 2-Dimensional PAGE M, Abandon scheme and start again 20K Store your material... 10K- 5K Using antibody to protein 10 Free versions of Protein Purification are available for the iPhone and Android phores. Protein Purification is now avalable for Windows (XPMista/7/8). Download Now It is also available for OS X 10.6 (Snow Leopard) and later. Download Now ur Moodle, Blackboard or other website, please consider making a donation to support the development, maintenance and delivery of Pra es, access via links from non-donor sites may be restricted or denied without notice.) MacBook Air 9 3 5 6

Answer 1

Protein purification- It is the process by which proteins are separated from a mixture of other complex individuals.

Some of the important methods of purification are-

1. ammonium sulfate fractionation

2. Heat treatment

3. Gel-Filtration

4. Ion-Exchange Chromatography

5. Hydrophobic interaction chromatography

6. Affinity Chromatography

Each of these is explained briefly below.

1. ammonium sulfate fractionation

the proteins are salted out, the concentration of salt at which protein precipitates changes for every individual protein. This property is used here to make the fraction of different proteins in the mixture.

2. Heat treatment

It can simply be stated as purification by denaturation.

3. Gel-Filtration

In this process, the size decides the purification. Here the sample is applied to a column of porous beads. depending on size they enter or retain the beads. According to pre-existing knowledge of size, and their entry into beads, purification is done.

4. Ion-Exchange Chromatography

In this process, beads are separated based on their charge. If the protein has an overall positive charge then it binds to a column of beads else not. Based on this they are separated.

5. Hydrophobic interaction chromatography

Here the separation is dependent on the vanderwaals force between proteins and ligands in the mixture. As different proteins have different associativity towards ligands, it makes to make and purify

6. Affinity Chromatography

Here the separation is based on the relatively high affinity of proteins with specific chemical groups.

Among all these methods, I would say Affinity Chromatography is the best of all methods because-

i. interaction highly specific.

ii. can isolate transcription factors also.

iii. effective to isolate the desired group.