Question

why is it important to use a small amount(1-5 mM) imidazole in the wash buffer for...

why is it important to use a small amount(1-5 mM) imidazole in the wash buffer for elution?

Homework Answers

Answer #1

peptides containing polyhistidine sequences can be easily eluted by either adjusting the pH of the column buffer or adding free imidazole to the column buffer.

A widely employed method utilizes immobilized metal-affinity chromatography (IMAC) to purify recombinant proteins containing a short affinity tag consisting of polyhistidine residues. IMAC is based on the interactions between a transition metal ion (Co2+, Ni2+, Cu2+, Zn2+) immobilized on a matrix and specific amino acid side chains

Know the answer?
Your Answer:

Post as a guest

Your Name:

What's your source?

Earn Coins

Coins can be redeemed for fabulous gifts.

Not the answer you're looking for?
Ask your own homework help question
Similar Questions
Miniprep DNA Wash Buffer (PE) contains: 10 mM Tris-HCl pH 7.5 and 80% ethanol (v/v) Answer...
Miniprep DNA Wash Buffer (PE) contains: 10 mM Tris-HCl pH 7.5 and 80% ethanol (v/v) Answer the following: a. You are asked to prepare 80 mL of DNA Wash Buffer (PE). How many mL of 100% ethanol should be added, then the total volume adjusted to the 80 mL final volume?
What is the reason of adding wash buffer twice while purifying DNA and why it is...
What is the reason of adding wash buffer twice while purifying DNA and why it is diluted with 100% Ethanol?
A procedure has 250 mM pH = 5 acetate buffer. 1) What concentrations of acetic acid...
A procedure has 250 mM pH = 5 acetate buffer. 1) What concentrations of acetic acid and sodium acetate should you use? 2) If the procedure calls for 4 L of the mentioned buffer solution, how many grams and moles of each buffer component will be needed?
To prepare 100 mL of 25 mM Tris buffer at pH 8.0, calculate the amount of...
To prepare 100 mL of 25 mM Tris buffer at pH 8.0, calculate the amount of Tris base and Tris·HCl needed to make the buffer. (Hint: find out the pKa of Tris·HCl and use the Henderson-Hasselbalch Equation.) Describe how do you want to make the solution, including how much water to add, how to adjust pH, etc.
Why is it not appropriate to use a small base amount in % analysis in evaluating...
Why is it not appropriate to use a small base amount in % analysis in evaluating financial performance over years?
The mean diameter of a spindle in a small motor is supposed to be 5 mm....
The mean diameter of a spindle in a small motor is supposed to be 5 mm. If the spindle is either too small or too large, the motor will not perform properly. The manufacturer measures the diameter in a sample of 60 motors to determine whether the mean diameter has moved away from the target. They found the mean diameter of the sample to be 4.951 mm. From past studies it is known that the population standard deviation is 0.2...
why is it less effective to wash an insoluble precipitate with 15 ml of water once,...
why is it less effective to wash an insoluble precipitate with 15 ml of water once, than it is to wash the precipitate with 3 ml of water 5 times See answers (1)
What is the law of small numbers theory? Why is it important?
What is the law of small numbers theory? Why is it important?
Why do you use distilled water to wash the organic layer and anhydrous sodium sulfate in...
Why do you use distilled water to wash the organic layer and anhydrous sodium sulfate in an extraction procedure
1.   What is the order of elution of: ethyl acetate (88.11 MM), acetic acid (60.05 g/mol),...
1.   What is the order of elution of: ethyl acetate (88.11 MM), acetic acid (60.05 g/mol), and dimethylamine (45.08 g/mol), off the following columns? The pKas of these are: 25, 4.75, 10.7, respectively, and you are using a neutral running buffer. a)   Reverse phase/hydrophobic    __________ then __________then __________ b)   Anion exchange       __________ then __________then __________ c)   Gel exclusion           __________ then __________then __________
ADVERTISEMENT
Need Online Homework Help?

Get Answers For Free
Most questions answered within 1 hours.

Ask a Question
ADVERTISEMENT