Gel Filtration Sephadex G 50

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  • เผยแพร่เมื่อ 24 ธ.ค. 2024

ความคิดเห็น • 88

  • @pantelitsadimitriou4166
    @pantelitsadimitriou4166 4 ปีที่แล้ว +3

    The most useful video on filtration columns!

  • @Ahmadkhan-bq2hc
    @Ahmadkhan-bq2hc 4 ปีที่แล้ว +6

    Awesome awesome .
    I was reading the textbook and not getting it .
    But after watching it whole stuff is clear .
    Thank you much ❤️ for this video.

  • @joshuarenfroe6548
    @joshuarenfroe6548 7 ปีที่แล้ว +4

    Very clear demonstration. Thank you

  • @muhammadafan5054
    @muhammadafan5054 3 หลายเดือนก่อน

    Well explained
    Great 👍👍

  • @AmirusSaleheen
    @AmirusSaleheen 7 ปีที่แล้ว +6

    Hi, this is a very helpful video. I had a question. What is the best way to tell which fraction contains the protein if the everything we are dealing with is colorless?

    • @acr92651
      @acr92651  7 ปีที่แล้ว +3

      Absorbance at 280 nm is commonly used to follow protein concentration. A protein assay (e.g. Bradford assay) can also be used to determine protein concentration.

    • @apurvakmr
      @apurvakmr 5 ปีที่แล้ว

      Uv spectrometry

  • @raquellaneiger3302
    @raquellaneiger3302 7 ปีที่แล้ว +15

    I absolutely love this video! Thank you!

  • @ekonglove4413
    @ekonglove4413 3 ปีที่แล้ว

    Thank you so much, I really did understand. Please is there another video for calculations as to help one plot a graph? If there is one, I will be waiting. Thank you once again

  • @DoctorCFlare
    @DoctorCFlare 5 ปีที่แล้ว +5

    How do you know when to swap the test tube drain caches?

    • @susannewalford5099
      @susannewalford5099 2 ปีที่แล้ว

      When the buffer is equilibrating the column, we count the number of drops it takes to get to 10 mL. Then we divide by 10 to get the number of drops for 1 mL. Students collect 1 mL aliquots into their tubes by counting the drops per tube.

  • @7infonews
    @7infonews 5 ปีที่แล้ว +1

    I take plant extract, i want different component of plant extract, which method is useful and accurate to separate plant extract component?

  • @reenamaedado107
    @reenamaedado107 3 ปีที่แล้ว +1

    would the steps be the same (+ the buffer used) for Sephadex LH20?

  • @michaelgimenez4032
    @michaelgimenez4032 4 ปีที่แล้ว +4

    If anything catches on fire at the lab tonight, i will blame you cause i will follow your method.

  • @christine2689
    @christine2689 4 ปีที่แล้ว

    Very nice video presentation!

  • @jemyjacob429
    @jemyjacob429 3 ปีที่แล้ว +1

    May l know the concentration of both the dyes .

    • @susannewalford5099
      @susannewalford5099 2 ปีที่แล้ว

      I was also interested in the concentration per dye.

  • @magdalenajaninalaska617
    @magdalenajaninalaska617 ปีที่แล้ว

    What is the column you are using? Thank you.

  • @tahrimasaihahuq6762
    @tahrimasaihahuq6762 หลายเดือนก่อน

    How much volume buffer did you use for the 9 g?

  • @poojashetve2669
    @poojashetve2669 3 ปีที่แล้ว

    Very nice demonstration 👏❤️..Thank you🙏

  • @purvibangoria8173
    @purvibangoria8173 3 ปีที่แล้ว

    Such a helpful demonstration.
    Could u help me with few queries..
    Does the flow rate of buffer hampers the separation of protein? What should be the flow rate and what amount of protein can we load into the resin?

  • @poojagujjer118
    @poojagujjer118 2 ปีที่แล้ว

    How to make blue dextran.. We should dissolve into distilled water or in phosphate buffer plz tell me

  • @rozgol2706
    @rozgol2706 4 ปีที่แล้ว +2

    Hello Dear 👋
    Thank you for good tutorials actually I have one question is there any different between Gel Filtration LH-20 and HiTrap Desalting because both packed with G-25 and for purification secondary metabolite I need LH-20 but HiTrap more convinced I will appreciate if you give me good answers because I am working on unknown components

  • @mel_from_hell
    @mel_from_hell 6 ปีที่แล้ว +3

    One of the best videos regarding size exclusion chromatography. I have some questions if you do not mind.
    I have bought Superdex 75 resin. If am not mistaken, I should pour the resin and let it for one or two days in a falcon tube eg to calculate the slurry (if I pour 10 mL of the resin and the new measurement is 5.5 mL it would be 55% of slurry). And then add this slurry into the column. My question is if I understood that correctly.
    Also, my column can host up to 16 mL, what is the preferred bed height to apply? I saw you have written about a 1:12 ratio of volumes.
    Finally, after packing a column, how many times can it be reused?
    Thank you in advance

  • @JoyceGem
    @JoyceGem 5 ปีที่แล้ว +1

    Good day, sir. Can I ask what kind pf column is that? Can I use the conventional glass open column (similar to those used for silica gel)?

  • @VelNasmr
    @VelNasmr ปีที่แล้ว

    s it possible to separate the sizes of DNA?

  • @isaacsnowhite104
    @isaacsnowhite104 4 ปีที่แล้ว

    Very nice demonstration..Thank you.

  • @manicpixiedreamgirl7930
    @manicpixiedreamgirl7930 6 ปีที่แล้ว +2

    This is a very useful explanation. Thanks.

  • @sciseeker-l9s
    @sciseeker-l9s ปีที่แล้ว

    Hi, I used sephadex G25 and packed the column similar to what's shown in the video. However, fluorescein doesn't retain as in the video but travels right though. Do you apply pressure in packing the column?

  • @jordannelewis9589
    @jordannelewis9589 6 ปีที่แล้ว

    Hello. My lab manual says to "Pour a column (Sephadex G-75 slurry) so that the bed is about 13 cm in height. Equilibrate the column in the elution buffer." Would equilibrating the column be what you did from 1:45 to 2:13?

    • @acr92651
      @acr92651  6 ปีที่แล้ว +1

      Equilibration means allowing buffer to flow through the column so that the entire gel bed is at a constant pH and ion concentration. Adding two or three column volumes of buffer to the top of the gel bed and letting it run through will ensure that the buffer concentration is uniform throughout.

    • @7infonews
      @7infonews 5 ปีที่แล้ว

      @@acr92651 I take plant extract, i want different component of plant extract, which method is useful and accurate to separate plant extract component?

  • @lmackenzie89
    @lmackenzie89 9 หลายเดือนก่อน

    Excellent video! Thank you!

  • @shaktichandramondal1923
    @shaktichandramondal1923 2 ปีที่แล้ว

    Can I use distilled water in replace of tris buffer?

  • @avneetkalra1145
    @avneetkalra1145 3 ปีที่แล้ว

    very nicely explained thank u so much!!!

  • @tarunrajput2951
    @tarunrajput2951 ปีที่แล้ว

    What's name of buffer

  • @29pete03
    @29pete03 4 หลายเดือนก่อน

    Thanks! This is incredibly helpful

  • @chelseyvandrisse7467
    @chelseyvandrisse7467 7 ปีที่แล้ว

    This was helpful, as there was no protocol on the GE website, you mention that 800 uL was a bit too much for this column, what volume of column per volume of sample is the limit?

    • @acr92651
      @acr92651  7 ปีที่แล้ว +1

      About 20 mL of gel bed for each 1 mL of loaded sample is a good ratio to use. In this video, the gel bed is about 10 mL for 0.80 mL of loaded sample (12 to 1 ratio)

  • @connordejager1125
    @connordejager1125 5 ปีที่แล้ว

    Why does the larger molecule move faster?

  • @reginaphalange1990
    @reginaphalange1990 3 ปีที่แล้ว

    What kind of column is that?

  • @jobayersonju766
    @jobayersonju766 5 ปีที่แล้ว

    Thanks for your nice video. What is the name of the empty column you used?

  • @franklindeoliveira6365
    @franklindeoliveira6365 2 ปีที่แล้ว +1

    what solvent did you put the 9 grams of sepahdex in? Was it water or alcohol?

  • @amanbhatia1860
    @amanbhatia1860 3 ปีที่แล้ว

    Can anyone help me to know: What's the scientific name of this type of tubes used for this purpose? Thanks

  • @AlOnEWolf007_JM
    @AlOnEWolf007_JM 3 ปีที่แล้ว

    How to extract ricin protein from this process??

  • @ailurophile4341
    @ailurophile4341 4 ปีที่แล้ว

    what if you drink it?

  • @BharathC-r3o
    @BharathC-r3o 7 หลายเดือนก่อน

    Thank you, you made my day 😊😊

  • @spencerrichards1045
    @spencerrichards1045 2 ปีที่แล้ว

    Can anybody recommend a process to purify an Oligonucleotide Mix in RNase Free Water. These oligos were only standard desalted. The volumes vary from 250mL to 1400mL.
    thank you
    Spencer

  • @brannigandupreez2083
    @brannigandupreez2083 5 ปีที่แล้ว

    I am using Sephadex LH20 to separate and purify proanthocyanidins from wine.
    Methanol is the solvent. Bed volume 290 ml, with 4ml/g of dry material used for given solvent. How big should my sample be without causing poor seperation/resolution?
    Thanks!

    • @JoyceGem
      @JoyceGem 5 ปีที่แล้ว

      The recommended sample volume would be 1-2% of the total bed volume according to the manufacturer.

  • @DocScoop7
    @DocScoop7 ปีที่แล้ว

    amazing

  • @sophiet5708
    @sophiet5708 7 ปีที่แล้ว

    Thank you this is a huge help! How much buffer did you swell 9g of sephadex in, in the beaker?

    • @acr92651
      @acr92651  7 ปีที่แล้ว +1

      About 20 mL of buffer is recommended for each gram of resin.

  • @vitokonte
    @vitokonte 2 ปีที่แล้ว

    thanks for tutorial!

  • @weibinding4487
    @weibinding4487 4 ปีที่แล้ว

    thanks for your help

  • @giorgosvlachopoulos1615
    @giorgosvlachopoulos1615 2 ปีที่แล้ว

    Thank you!!!

  • @HalannaR
    @HalannaR 6 ปีที่แล้ว

    My gel (Sephadex G-100) keep running through out the columm. I dont know if it is the consistency or maybe the buffer I used to swell it up (sodium acetate 50mM pH 5,0 - the enzime Im looking to purify is well adapted to the this buffer) is not appropriate for it. Should I plug a small piece of cotton to the botton of my columm and then add the sephadex to avoid the gel to leak out?

    • @acr92651
      @acr92651  6 ปีที่แล้ว

      If the gel leaks out of the column, you will need to add something like cotton or glass wool to keep the beads in. The columns I use have built-in frits that retain the fine gel beads.

    • @amanbhatia1860
      @amanbhatia1860 3 ปีที่แล้ว

      @@acr92651 Can you please let me know the scientific name of the column tubes you have used. I would like to purchase the same.. Thanks

  • @sabatavana5127
    @sabatavana5127 3 ปีที่แล้ว

    Thank you!

  • @StockMarketTrading555
    @StockMarketTrading555 6 ปีที่แล้ว

    Can you please guide me how I can purify a protein of mw 40kd using chromatography from a fermented broth.

  • @swarnaliartwork
    @swarnaliartwork 4 ปีที่แล้ว

    Nicely explained

  • @gioso667
    @gioso667 7 ปีที่แล้ว

    What's the expiry date of the packed gel in the column?

    • @acr92651
      @acr92651  7 ปีที่แล้ว

      When refrigerated, the gel can last at least a week, maybe a few weeks. If refrigerated with an azide-containing buffer, the gel can last for a year or so (0.1% sodium azide).

  • @sugasmile8122
    @sugasmile8122 4 ปีที่แล้ว +1

    Thank you so much for this 😄😄😄

  • @alhaeri1
    @alhaeri1 3 ปีที่แล้ว

    thank you

  • @achanyasjayan9195
    @achanyasjayan9195 3 ปีที่แล้ว

    Thankuuuu

  • @noobnoob8922
    @noobnoob8922 ปีที่แล้ว

    Neeru ma'am bought me here.

  • @DJmisterpeluca
    @DJmisterpeluca 3 ปีที่แล้ว

    Nice

  • @lucasfohn
    @lucasfohn 7 ปีที่แล้ว

    Thank you so much for making this video

  • @doupleyy5684
    @doupleyy5684 4 ปีที่แล้ว +1

    dank goethe uni so viele views

  • @sheikhaaqib6629
    @sheikhaaqib6629 4 ปีที่แล้ว

    Lovely

  • @sallyhungwe220
    @sallyhungwe220 4 ปีที่แล้ว

    Thank you sooo muchhh❤️❤️❤️

  • @helen5930
    @helen5930 7 ปีที่แล้ว

    what was the buffer you used?

    • @acr92651
      @acr92651  7 ปีที่แล้ว +2

      30 mM Tris, pH 8.0

  • @antoniojafar1197
    @antoniojafar1197 4 ปีที่แล้ว

    Thank you, :)

  • @kartikmandal455
    @kartikmandal455 3 ปีที่แล้ว

    Its very useful.

  • @deninta8387
    @deninta8387 7 ปีที่แล้ว +2

    This vid is very help, thank you so much!

  • @ruguoserliegise2716
    @ruguoserliegise2716 4 ปีที่แล้ว +1

    Biochem squad

  • @levelone7511
    @levelone7511 7 ปีที่แล้ว

    Thank u sir

  • @jatin1995
    @jatin1995 5 ปีที่แล้ว

    useful

  • @HStiglitzz
    @HStiglitzz 7 ปีที่แล้ว

    nice, it will help :)

  • @halikarnak1862
    @halikarnak1862 2 ปีที่แล้ว

    sigma sephadex

  • @NaniSenpaii
    @NaniSenpaii 5 ปีที่แล้ว

    To many bubbles!
    But still a nice video ✌

  • @Sammythakidd
    @Sammythakidd 4 ปีที่แล้ว +3

    wer ist hier wegen dem Biochemie Praktikum hahah?

  • @niranjancv5937
    @niranjancv5937 4 ปีที่แล้ว +1

    Can you please guide me how can I purify a protein of MW 20-25 KDa using chromatography from a conjugation buffer