$20 Bonus + 25% OFF +
Securing Higher Grades Costing Your Pocket? Book Your Assignment at The Lowest Price Now!
Aiden Keith Student Reviews Student Reviews Student Reviews Student Reviews Student Reviews 4.6/5 hire me


Describe about the Chromatographic Purification?


Purification of Soluble His-Tagged Proteins

The ClpA is the protein of our interest is an ATP dependend chaperon and regulatory component of ClpAP protein basically expressed in E.coli cells. This expression of recombinant protein could be analysed by presence of small size of amino acids i.e. 6 histidine tag which will be conjugated to its C or N terminus of protein. It can purify by using metal chelate affinity chromatography method having concept based on strongly binding affinity of Nickle (II)-nitriloteriacetic acid (Ni-NTA) and NTA act as chelating ligand. The 6 histidine ligand binds with Ni2+ ion to NTA through four ligand sites leaving two sites vacant for binding with 6 his- tag (Zhao et al, 2010). The strong affinity between Ni conjugate NTA with hexahistadine tag lowers the chances of unspecific binding of protein in Ni-NTA beads. The washing of column allows the highest affinity binding of hexahistadine tag protein with IMAC resin by nearly neutral condition buffer.  The next step to elute the tagged protein, in this step the His tag protein is eluted by elution buffer twice with different concentration. The elution should be done in concentration gradient where the first concentration should be low which remove the unspecific binding of protein and the second elution should be done with high concentration that is used to remove the hexahistadine tag protein from the Ni-NTA (Crowe et al, 1994).  Finally, the prurity of isolated pure his-tagged protein will be identified using techniques like SDS-PAGE and 2-dimesion gel electrophoresis leading to identification of pure protein.



Zhao X, Li G, Liang S (2013) Several Affinity Tags Commonly Used in Chromatographic Purification. J Anal methods Chem: doi: 10.1155/2013/581093

Crowe J, Dobeli H ,Gentz R, Hochuli E ,Stibber D, Henco K (1994) 6xffis-Ni-NTA Chromatography as a Superior Technique in Recombinant Protein Expression/Purification .Methods in Molecular Biology .Mol biotechnol 31:371-387


MyAssignmenthelp.com is one of the noted service providers that deliver essay help. We provide tailored essay assistance to make sure that student gets online essay help exactly in the way they want it to be written. We at MyAssigemnthelp.com have built teams of consultants, who readily attend every query related to help me writing my essay. We provide essay writing help in forms of tips and steps in order o assist students with tough essay assignments.

Most Downloaded Sample Of Management

Toulin Method Of Argumentation

Download : 289 | Page : 1

You are required to write a researched argument essay that convinces persuades the reader of your position / stance. This is an academic, researched and referenced document that demonstrates your understanding of the objectives of the course. The structure of the argument will follow the TOULMIN METHOD of argumentation.

Read More arrow Tags: Australia Arlington Management University of New South Wales 

Consumer Behavior Assignment

Download : 211 | Pages : 9

Executive Summary The purpose of this report is to elaborate the factors which are considered by individuals before selecting an occupation. Choosing an occupation is indeed a major decision of everyone’s life and it does give a shape to where he would be heading in the course if time. The report begins with a list of various occupations which have been going since the human civilization started though it has indeed changed its form to...

Read More arrow Tags: Australia Arlington Management University of New South Wales 

Internet Marketing Plan For River Island

Download : 371 | Pages : 13

Introduction With the increase enhancement in the field of technology, it has been considered essential by the businesses to implement such technology in their business processes. Further, the increasing demand from the customers regarding the better quality of products and services delivered to their doorstep has also necessitated for the businesses to implement the e-business strategy in their business processes. E-business which is common...

Read More arrow Tags: Australia Arlington Management University of New South Wales 

Strategic Role Of HR In Mergers & Acquisitions

Download : 330 | Pages : 9

Executive Summary In a merger & acquisition, role of an HR has emerged as a very critical function. At each stage of merger and acquisition process, HR plays a strategic role. The importance of role of HR part in integration of the merged companies should not be taken lightly and should be handled very carefully and with due planning. At pre acquisition stage, due diligence should be taken before the start of the process and HR must carve ou...

Read More arrow Tags: Australia Arlington Management University of New South Wales 

Relationship Between Knowledge Management, Organization Learning And HRM

Download : 362 | Pages : 7

Introduction In this competitive business environment where every business organization is trying to attract the customers of each other, it becomes essential for these organizations to remain competitive by innovating new ideas and thoughts, so that value added products and services can be provided to the customers. The motive of such organization in providing value added services to their customers, require their employees to be quite effect...

Read More arrow Tags: Australia Arlington Management University of New South Wales 
Free plagiarism check online Free plagiarism check online on mobile
Have any Query?