A full and complete explanation about western blotting technique. Watch also SDS-PAGE: • The principle of SDS P... Southern and Northern Blotting Techniques: • The Principle of South...
Пікірлер: 104
@ciao_abhi6 жыл бұрын
I cannot stress how helpful this is ... no other video explain this process with this much clarity. Definitely helping me on MCAT test
@sgdrifter3 жыл бұрын
Hi Dr, you explained everything so clearly! I have watched nearly every video you posted! Thank you so much!
@DA-sj2gw6 жыл бұрын
Thank you for a this great explination! Im going to use this technique in my bachelors thesis and this video clarified a lot of things for me!
@WS-xc4yp5 жыл бұрын
I havent finished watching this video yet and boy you are doing a great job. I have been looking for someone who explains the protocol and the theory behind it and this is exactly what i need. Please keep posting 👍👍👍😘😘😘😘
@joelarnoldngassa87202 жыл бұрын
She's a girl
@97alvarezn6 жыл бұрын
Thank you very much, I'm a med student preparing for my Microbiology exam and this helped a lot!! :))
@kamranheraji55532 жыл бұрын
The videos on this channel are great and helped me a lot to better understand the techniques. Your teaching method is excellent, and it is very useful to explain the reason for using each substance and solution. I hope you continue this great videos. Thank you so much!
@amoon40367 жыл бұрын
first time I understand it .. thanks millions.
@baselbasels33217 жыл бұрын
thank you for your easily explanation it's help me very well ... please don't stop these interesting videos
@augustoli51003 жыл бұрын
well explained!! the graphs are really clear to understand !!
@jackbauer78494 жыл бұрын
Very well explained and extremely helpful, thanks so much!
@julymgreenday19936 жыл бұрын
THANK YOU!!! ♥ You helped me a lot. Greetings from Argentina (:
@mg21722 жыл бұрын
This was explained so well. Thank you so much!
@hrushikeshhrushi37403 жыл бұрын
Thank you madam. Your videos are making the concepts very easier
@naqibrahimi99584 жыл бұрын
Thank you very much for this informative and detailed video. I enjoyed it very much.
@esraahader46045 жыл бұрын
Thank you very much... You are explanes amazingly.... Pleas complete this video
@fieldtripinmyparty5 жыл бұрын
you're simply ... Professional
@dawielslawischki37564 жыл бұрын
Thank you for this informative video, finally I'm able to understand WB :)
@colin-kun36113 жыл бұрын
Awesome video.. it helped me a lot to understand the principle of western blotting.
@romelsoyza41606 жыл бұрын
How do we determine the precise Time and Voltage required for our protein of interest? Can we look it up somewhere, or is this something that requires some trial and error? Also, if for example, we have 10 different proteins of interest that we are looking for in our samples, does each protein have their own optimal Time and Voltage (I am assuming yes)? If yes, how can we optimize the Time and Voltage so that we get ALL of our 10 proteins of interest on to the membrane without having some of them being left behind in the gel or in the filter paper?
@multiplEYE16 жыл бұрын
Thank you so much for the video, helped me a lot!!! Appreciate it.Greetings from an italian student ;-)
@biomedicalandbiologicalsci49896 жыл бұрын
This makes me happy .. thank you :)
@sukieyakie4 жыл бұрын
ugh so good thanks for enlightening me on western blotting
@sharbaridas49125 жыл бұрын
Very helpful video!!! Thanks so much
@szechungyau7356 жыл бұрын
Very useful video thank you so much!
@aeorherhe67994 жыл бұрын
Thank you. This was very helpful.
@AD-dy1hi2 жыл бұрын
Simply amazing! Thank you 🙏
@faisaljj543ztrhf6 жыл бұрын
Very informative. Thank you
@ranveerpratapsingh16836 жыл бұрын
thanks for such an explainatory video
@meissaabrahao74323 жыл бұрын
thank you so much for your explanation. Very nice
@krisztiankatona87834 жыл бұрын
I have a question regarding the secondary AB. if the first Ab is mouse anti-human why isnt the secondary let’s say goat anti-mouse? Isnt it a mistake? Thanks
@izzatiawgrahman33975 жыл бұрын
thankyou for the explanation... it is easier to understand :)
@alaaobeid43284 жыл бұрын
Really Amazing There Is video about native electrophoresis plz?
@ranahaddad54462 жыл бұрын
You are amazing, thank you so much!
@dancanrogers95124 жыл бұрын
great work and well explained
@pooryaforoutan66293 жыл бұрын
so clear simple and useful thanks
@rehanaakhter91933 жыл бұрын
well explained in a simplified way
@niveditasharma93143 жыл бұрын
Could to please made a video for developing a blot in chem doc which consists of dos n donts
@nshimirimanajonas10772 жыл бұрын
Thank you. powerful video
@quanganhpham91394 жыл бұрын
OK. So I have questions. 1) Is there a "ladder" for SDS-PAGE? Otherwise I don't see how can I determine specific molecular weight numbers 2) SDS denatures proteins, would that interfere with the primary antibodies from recognizing the protein of interest given that it is denatured? 3) The "sandwich" is placed in PLASTIC container. Why use plastic, a non-conductor, when we are going to electrically push the proteins? Does the transfer buffer gets into the plastic bag, or stay completely outside?
@Leandier976 жыл бұрын
Short question.: Why do all proteins move to the anode if you prevent SDS-binding with methanol? Whithout SDS-binding some basic/alkaline proteins would be positively charged, wouldnt they?
@biomedicalandbiologicalsci49896 жыл бұрын
Hei, proteins here are already bound to SDS, because as I said; before applying the western blotting technique we applied the SDS-PAGE in which all the proteins were bound to SDS. So the Methanol is added to the transfer buffer in only 20%, so during the run, methanol will lightly detach the SDS from the proteins (not completely) in order to enhace their ability to bind to the membrane.
@asmasomadayo24 жыл бұрын
finally i understand WB, good bless u 😭🌹🌹🌹🌹🌹🌹🌹🌹🌹🌹🌹🌹🌹
@skyimran65724 жыл бұрын
Nice explanation mam...likes your videos
@tamirissociareli64684 жыл бұрын
Amazing video!
@icecreamgirl2422 жыл бұрын
"...from the mouse or from the rat or from the rabbit or whatever!" 😀 Anyway, I'm moving from SDS-PAGE to Western Blotting in my research lab, and this video was very informative, helping me understand the technique and protocol!
@SDscienceproductions3 жыл бұрын
Perfect explanation thank youuu
@yuvalgal-shahaf27822 жыл бұрын
great explanation
@zienanema46913 жыл бұрын
You are very very very wonderful thank you very much teacher
@mammaamanda12775 жыл бұрын
omg finally i understand this!!!
@akhtar19974 жыл бұрын
This is awesome! Thanks! :)
@towfiquebhuiyan45154 жыл бұрын
Such a nice presentation..😘😘
@dianas98196 жыл бұрын
Thank you!
@minseon2133 жыл бұрын
Thank you so much.
@alexei8468 Жыл бұрын
Muchas gracias!
@mohameda.hassany65624 жыл бұрын
you are amazing thank you so much
@nadeekadimuthu90075 жыл бұрын
Hi can I please know what is the reason for using glycine ? is it like a driver molecule for the proteins to separate ?
@julianomachado70503 жыл бұрын
I am also interested in this potential explanation!
@andrewayman31696 жыл бұрын
Really you are very good .... Actually this video helped me 😄
@Biomeducated5 жыл бұрын
Do you have a control protein blotted as well? I assume your proteins of interest are from a cell lysate, no? A housekeeping protein like GADPH, or something like Actin? Or some bacterial protein if you have expressed the poteins to have them purified later on? If the band size/intensity of the Controls is the same in each condition, then you indeed have expression differences between wild-type and mutant.
@julianomachado70503 жыл бұрын
Has ethanol the same principle as methanol? We use a commercial "turbo buffer" which is recommended to use ethanol instead methanol!
@NidhiPatel-er2su7 жыл бұрын
I have a question... I understand that the primary Ab is extracted from something other than a human (such as a mouse or rabbit). I also understand that the secondary antibody is made anti- whatever the species used to make the primary antibody. However, how is the secondary antibody produced? For instance, if you have a human protein, a mouse primary antibody, then would you need to synthesis the secondary antibody in another animal such as a rabbit? What happens if you synthesis the secondary antibody in a human or mouse?
@biomedicalandbiologicalsci49897 жыл бұрын
Hello ... The secondary antibody is an IgG that has a specific affinity to the proteins of a certain species such as mouse or rabbit or sheep. It is also produced in an animal by hyper-immunizing (injecting a high concentration of immunoglobulins) the animal with purified immunoglobulin fractions from a normal mouse (or any other animal) serum to produce High-affinity antibodies. Yes, usually the secondary Ab is produced in another animal because you cannot produce anti-mouse secondary Ab in a mouse because if you inject mouse immunoglobulins into a mouse then the mouse will not produce Ab against them. On the other hand, you can produce anti-mouse Ab in humans but usually, we dont produce Ab in humans (ethically). However, if you are studying sheep proteins for example and you use mouse primary Ab you better not use sheep secondary Ab to prevent unspecific binding. So usually for example, if we are studying human proteins we use mouse anti-human primary Ab and rabbit anti-mouse secondary Ab.
@devashishkumar57424 жыл бұрын
You told methanol removes SDS from protein, then how it is still negatively charged?
@arimene37483 жыл бұрын
Thank you so much
@meravya16 жыл бұрын
how can i know how much time and voltage i need to my proteins? it depends on what? how can i know if my proteins run away from the membrane or some stayed on the gel?
@Biomeducated5 жыл бұрын
If you have included your 'protein ladder/standard' in SDS-PAGE (as you should! ;)), then you will see it on your blot (the colours of the ladder are seen)...if not, well, then your samples have migrated into the solution and you can start all over again :p point is, you cannot see it during the run. Make sure you get it right immediately, and stick to your own routine, always, when preparing the cassette! Time and voltage depends. Anyway, you should be careful with higher voltages, as this might increase temperature, and maybe decrease the quality and resolution of your blot. As I remember, we used 25V for 'overnight' (ca. 16h) (and put the whole setup in an ice filled bin!), if you start blotting at the end of the day, OR 45V for intraday (ca. 4h). This worked fine, but ideally you should optimize for your own equipment yourself :)
@DA-sj2gw6 жыл бұрын
I have a preformed a WB and im comparing a wildtype and mutant protein and the results show that we have more intensity in the mutant samples than the wildtype samples. The only thing this tells me is that we have more of the protein of interest in the mutant compared to the wildtype right i.e. it doenst give me any other information?
@Biomeducated5 жыл бұрын
Do you have a control protein blotted as well? I assume your proteins of interest are from a cell lysate, no? A housekeeping protein like GADPH, or something like Actin? Or some bacterial protein if you have expressed the poteins to have them purified later on? If the band size/intensity of the Controls is the same in each condition, then you indeed have expression differences between wild-type and mutant.
@hfyyshnur_3 жыл бұрын
THANK YOU SO MUCH
@imtiazahmad56315 жыл бұрын
Please do upload videos on biological technique
@hussaino69636 жыл бұрын
Fentastic !!! Thanks a lot..
@biomedicalandbiologicalsci49896 жыл бұрын
You are welcome :)
@user-tp8tu7ny6v5 жыл бұрын
Can anyone explain that why proteins are negatively charged to me?
@amir.sajadjafari41266 жыл бұрын
it was very helpfull
@sonalisahoo32053 жыл бұрын
Very much thankful ☺️
@hitkarshkushwaha24346 жыл бұрын
THANK YOU MA'AM ! PLEASE UPLOAD VIDEO ON HPLC
@biomedicalandbiologicalsci49896 жыл бұрын
kzbin.info/www/bejne/oGfXYY1jq65podk go to this link .. and subscribe the channel so you can see all the new videos :)
@ajaysubbaroyan24614 жыл бұрын
Thank you !!
@nawrozsabah23793 жыл бұрын
Thank you ❤️❤️❤️
@kundlikrathod88415 жыл бұрын
thank you !
@murtadhabasheer16055 жыл бұрын
Thank you a lot
@luisalbertomercadovianco23733 жыл бұрын
Excelente!
@samarthsharma68103 жыл бұрын
awesome
@ithirstyforknowledge5 жыл бұрын
Thanks
@TuanAnhNguyen-gr4bk4 жыл бұрын
What 's the filter papers and sponges for?
@bhagyashreejadhav87574 жыл бұрын
To compress nitrocellulose membrane tightly on gel..
@mahmoudtolba88327 жыл бұрын
so great ..go on
@biomedicalandbiologicalsci49897 жыл бұрын
Thank you ... stay around many interesting videos are coming up :)
@chicong876 жыл бұрын
Thanks a lot, :)
@biomedicalandbiologicalsci49896 жыл бұрын
Welcome :)
@vanyabawa64402 жыл бұрын
Very I formative
@deepakthakur96415 жыл бұрын
Good
@vinayakca8925 жыл бұрын
You are too good mam
@sukantamandal81064 жыл бұрын
So helpful more details and immunofluorescence,imuunochemistry,immunodiffusion,emsa,