The Principle Of Ion Exchange Chromatography, A Full Explanation

[@ml244716]

Another rule, is that you should use a pH at least 1 unit away from the PI in order to make sure the protein/peptide is completely protonated/deprotonated. So, if the PI is 2, and the pH is 2.3, you will probably not get good retention of your target peptide.

[@ruqaiyatasneem8594]

It makes your protein more soluble in the buffer

[@IRRevs]

if the ph of the medium is lower than the pI of the protein, then the protein will be protonated and hence will be positively charged. If the ph is higher than pI then less proton are available in the medium and so the protein will lose its hydrogens from the side chains and become negatively charged.

[@piyushsen7502]

Ya you are right

[@einsteinyamat3894]

For someone who has not enrolled in a Biochemistry laboratory class, this has helped me visualize what to do in the future. Thank you.

[@AsYouWishEquestrian]

Thank you for these great videos! My biotech LAB classes are only virtual right now (which is stupid) and we are not learning how these techniques actually work, but with your videos I will be ready to get a job.

[@HIGHER7RUTH]

You can work for me where you from?

[@reemmahmoud6342]

Thank you you explained that 2 substances are in ion column...subs A directly attached to resin and subs B that is bound to it by an ionic bond and that B is the one to be exchanged with the analyte...thank you for revealing this

[@harasama7738]

you have no idea how you SAVED ME , much much love for you my savior

[@ajithprasad13]

Very nice presentation and highly useful...

[@milkasvilar8936]

Wow!!! You are so awesome! This is the best video that explained this method so detailed and so understandable! This was so helpful! Thank you!

[@manishasaboo9445]

Your videos are amazing..... please keep making such videos and try to make videos frequently....videos are very helpful

[@patriciabarkoci2833]

very good explanation! definitely helped me to prepare for my upcoming exams . thank you for this. keep this up, you're doing great job

[@mohammadabouhassan780]

much thanks for the clear illustration! One more question I have is that how the exchange occurs between the two elements of same charges ( B and C) in other way what is the reason behind the settlement of the protein of interest (C) that we want to eliminate and having the exchanged element (B) in our outflow??

[@reemmahmoud6342]

It can be because subs A has more affinity to C than B...or because the concentration of C is much higher than B

[@rredding]

Thank you for your clear explanation! 😊

[@joyjoy9490]

It was beautiful I understood everything Great job 👏

[@chandrusekar4644]

It's really helpful for me, add some other's techniques like HPLC, GC etc...

[@riddhimanchatterjee4195]

Great video. Thanks a lot🎉.

[@Alman_und_Javid]

Thank you very much for explaining so good.

[@Linges-dm8dw]

Super explaination

[@majdaelhassani4145]

It was really helpful, I have only one observation is that you talk aloud sometimes. But the explaination was clear and great for me. Thank you !

[@biomedicalandbiologicalsci4989]

Thank you for the remark ... trier to solve the problem in my two new videos :)

[@mohdhisyamuddin1016]

Thank you for the effort. Keep it up!

[@meenakshichouhan4147]

Excellent

[@paulinamichaud4580]

When do you use Ion or Absorption? Will the choice be subject to the substance of study?

[@nikunjkumarkanani9018]

It's best video for this topic

[@rikkigupta9546]

Thanks a lot Mam very informative lecture

[@sarabio5269]

please in ion chromotograph Which columns are the most preferred in this technique

[@ananthiponnusamy7938]

@ 11.15 if pH is less, than pI, it would be positive charge

[@biomedicalandbiologicalsci4989]

When the Ph of the medium is less than the PI of the protein .. than the protein is definitely positively charged

[@maggiejameel6725]

Thank you for the great and clear explanation

[@midosala8049]

Great Video! Explained really well! Thank you so much

[@mentawai3959]

very well explained

[@Grandpierrefull]

Video is great thank you ! Tho, for the last part, the Salting out is not so clear. I didn't undersand why proteins with 1 nucleotide are eluted first.

[@Adaeze_Nduka]

I’ll probably think it’s because it has less binding groups attached to the stationary phase coupled with its having the lowest molecular weight.

[@ruqaiyatasneem8594]

How will B+ on anion exchange resin get replaced by our protein of interest?

[@ithirstyforknowledge]

you probably said reverse at 11:15

[@drjahanzeb1135]

Very informative video. How I can digest environmental samples to run it on IC ?

[@deemamuhaisen2981]

You're the BEST

[@paulinamichaud4580]

What defines what is Polar or non-polar?

[@aratimaurya8464]

how to determine Ph of protein my protein has PI = 5.51 , what will be it's ph at 7.2 buffer .

[@All_movies_hub_326]

Maam if ph of sol is less than 5 for pi5 then protein should be +vly charged here is confusion

[@ithirstyforknowledge]

yes. so where is the confusion?

[@All_movies_hub_326]

You have asked protein is -vly charged at ph less than 5 if pi of protein is 5

[@user-hx1ho6tl1b]

this really helped a lot! thank you !

[@suonnneee3396]

It’s really helpful, but isn’t it “anion” not “inion”?

[@zeinabgh5989]

thank you so much, it helped me a lot!

[@egs8545]

very helpful

[@nizamuhd8717]

is there any differences of chromatogram for cation and anion ion exchange chromatography?

[@biomedicalandbiologicalsci4989]

No difference in the chromatogram for inion and cation exchange chromatography ..

[@gauravresearch8]

@@biomedicalandbiologicalsci4989 Hi. Thanks for the wonderful video. But at place Anion has been written as Inion.

[@ramaprasad1028]

If pka is 5...then what method should be used for chromatography and what ph should be used for elution

[@chrischukwuma6469]

I would use anion exchanger( positively charged stationary phase), adjust my medium PH above the pka to deprotonate and make my Protein negative, I wouldn't want a lower pH that will make my Protein positive as there are higher chances of having other Proteins that are stable around that pH that can bind alongside my Protein. For elution, I will gradually decrease the inonic strength of my Buffer.

[@thanworker3160]

Thankyou so much

[@goodhealthaboveall5844]

Wonderful!!!

[@ghenwaismail7070]

what might be substance B?

[@ntinadov1477]

THANK YOU SO MUCH

[@ganeshaher5437]

Mam why we use Isopropyl alcohol or any organic modifier in Elution buffers during collection of Elute ( intereste protein) Actually I am using DEAE sepharoge fast flow resine for separate of lys-arg insulin from mixture. Here I am using 20mM Tris HCl+85mM Nacl+30%IPA? So why we use this IPA?

[@wumbeiyakubu2831]

what reaction takes place in this separations

[@ithirstyforknowledge]

ion exchange

[@einsteinyamat3894]

Have you heard of "Single-displacement reaction?"

[@taimouranjum654]

i want to test bromate by my drinking water then how i do it?

[@biomedicalandbiologicalsci4989]

You need first to isolate bromate from drinking water using anion exchange chromatography, then you elute bromate using salting out. And then you can detect the presence of Bromate using a type of spectroscopy or UV light detector. There are machines nowadays that combine all these steps together, they are specially designed to detect bromate in drinking water.

[@saneelakhan3081]

@taimour anjum why u want to do that test?

[@piyushsen7502]

Thanks

[@cokieeeeeeee]

Inion exchange chromatography? Check your slides before sharing them....

[@zygaf6252]

7:50 totally wrong explanation. It is not true that in pI half of all molecules of aminoacid is in cationic form and half of all molecules is in anionic form.

[@SayaSergeiGustave]

It is great but please when you talk can you step wiggling your mouth.

[@minasoltani5762]

So bad 😔

[@goodhealthaboveall5844]

Wonderful!!!