country
$20 Bonus + 25% OFF
Securing Higher Grades Costing Your Pocket? Book Your Assignment at The Lowest Price Now!

Question:

Describe about the Chromatographic Purification?
 
 

Answer:

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.

 

References:

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

OR

Cite This Work

To export a reference to this article please select a referencing stye below:

My Assignment Help. (2016). Chromatographic Purification. Retrieved from https://myassignmenthelp.com/free-samples/chromatographic-purification.

"Chromatographic Purification." My Assignment Help, 2016, https://myassignmenthelp.com/free-samples/chromatographic-purification.

My Assignment Help (2016) Chromatographic Purification [Online]. Available from: https://myassignmenthelp.com/free-samples/chromatographic-purification
[Accessed 21 February 2020].

My Assignment Help. 'Chromatographic Purification' (My Assignment Help, 2016) <https://myassignmenthelp.com/free-samples/chromatographic-purification> accessed 21 February 2020.

My Assignment Help. Chromatographic Purification [Internet]. My Assignment Help. 2016 [cited 21 February 2020]. Available from: https://myassignmenthelp.com/free-samples/chromatographic-purification.


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.

Latest Management Samples

BUSN20016 Research In Business 23

Download : 0 | Pages : 4

Answer: Problem statement Number of radical, significant and unexpected events have been seen in many industries in 21st century which have even impacted tourist activities in major countries like Australia (Beesley, Lisa, Davidson, & Michael, 2013). Global climate and diversified people culture need tourism companies embrace disciplined and continuous reformation to monitor future trends and problems to make uniform policies an...

Read More arrow

BUSN20016 Research In Business 22

Download : 0 | Pages : 3
  • Course Code: BUSN20016
  • University: Central Queensland University
  • Country: Australia

Answer: Problem statement In the apparel industry, Nike occupies the top most position. According to the reporting of the Wall Street Journal, it has been noticed that Nike shoes are losing its style. This is mainly due to the intense competition, which aggravates the complexities towards serving quality products to the customers. As a matter of specification, investors dealing with the basketball shoes have declined largely (Nike.com, 2018). T...

Read More arrow

BUSN20016 Research In Business 21

Download : 0 | Pages : 5
  • Course Code: BUSN20016
  • University: Central Queensland University
  • Country: Australia

Answer: Problem statement: For a prolonged period of time, developing and sustaining brand loyalty are two of the highly crucial factors of market research (Erdo?mu? & Cicek, 2014). Market research utilizes various tools to monitor and maintain the brand loyalty in the market segment. With the emergence of internet, social media marketing have huge popularity as a mean to maintain brand loyalty of the consumers (Laroche, Habibi & Rich...

Read More arrow

BUSN20016 Research In Business 20

Download : 0 | Pages : 4

Answer: Problem statement Brand awareness is an important aspect that impact’s consumer’s perception towards buying products and services (Jeon, 2017). Brand achievement arises when companies understands and oversees loyalty and brand image to implement strong impression among customers for making purchase decisions ( Carpenter, Bauer, & Erdogan, 2012). This theory concentrates on significance of brand image on sales...

Read More arrow

BUSM4687 Managing International Business Responsibly Findings

Download : 0 | Pages : 13
  • Course Code: BUSM4687
  • University: Royal Melbourne Institute Of Technology
  • Country: Australia

Answer: Introduction Today Corporate Social Responsibility (CSR) has become a very popular topic in the global market. With the increase of demand for corporate citizenship and transparency, CSR has been majorly required to embrace ethical, economic, social, and also environmental considerations. Through the current globalization changes and approaches, companies are expected to not only be of commercial value but also good corporate citizens...

Read More arrow
Next
watch

Save Time & improve Grades

Just share your requirements and get customized solutions on time.

question
We will use e-mail only for:

arrow Communication regarding your orders

arrow To send you invoices, and other billing info

arrow To provide you with information of offers and other benefits

1,202,444

Orders

4.9/5

Overall Rating

5,061

Experts

Our Amazing Features

delivery

On Time Delivery

Our writers make sure that all orders are submitted, prior to the deadline.

work

Plagiarism Free Work

Using reliable plagiarism detection software, Turnitin.com.We only provide customized 100 percent original papers.

time

24 X 7 Live Help

Feel free to contact our assignment writing services any time via phone, email or live chat.

subject

Services For All Subjects

Our writers can provide you professional writing assistance on any subject at any level.

price

Best Price Guarantee

Our best price guarantee ensures that the features we offer cannot be matched by any of the competitors.

Our Experts

Assignment writing guide
student rating student rating student rating student rating student rating 5/5

1692 Order Completed

98% Response Time

Alfred Dodd

PhD in Computer and Information Science with specialization in Database

Wellington, New Zealand

Hire Me
Assignment writing guide
student rating student rating student rating student rating student rating 5/5

2115 Order Completed

97% Response Time

Kimberley Chen

MPA in Accounting

Singapore, Singapore

Hire Me
Assignment writing guide
student rating student rating student rating student rating student rating 5/5

529 Order Completed

95% Response Time

Ivan Blank

PhD in Functional Human Biology

Wellington, New Zealand

Hire Me
Assignment writing guide
student rating student rating student rating student rating student rating 5/5

1265 Order Completed

97% Response Time

James Cook

Masters in Management

Wellington, New Zealand

Hire Me

FREE Tools

plagiarism

Plagiarism Checker

Get all your documents checked for plagiarism or duplicacy with us.

essay

Essay Typer

Get different kinds of essays typed in minutes with clicks.

edit

GPA Calculator

Calculate your semester grades and cumulative GPa with our GPA Calculator.

referencing

Chemical Equation Balancer

Balance any chemical equation in minutes just by entering the formula.

calculator

Word Counter & Page Calculator

Calculate the number of words and number of pages of all your academic documents.

Refer Just 5 Friends to Earn More than $2000

Check your estimated earning as per your ability

1

1

1

Your Approx Earning

Live Review

Our Mission Client Satisfaction

I have been submitting few assignments to this platform. They are very professionals, very happy with the results.

flag

User Id: 355073 - 21 Feb 2020

Australia

student rating student rating student rating student rating student rating

So happy to have found this service! I was in a pinch and they were able to assist in just a few hours!

flag

User Id: 376804 - 21 Feb 2020

Australia

student rating student rating student rating student rating student rating

hi my expert i was not aware this review was pending however i am please to tell you that i got an A grade so thank again cant wait to book your services again

flag

User Id: 270782 - 21 Feb 2020

Australia

student rating student rating student rating student rating student rating

excellent work! as always very pleased with my assignments and the timing of receiving it back.

flag

User Id: 237278 - 21 Feb 2020

Australia

student rating student rating student rating student rating student rating
callback request mobile
Have any Query?