I only watched this because of my assignment and gosh I've never been so overwhelmed by such laborious procedures!

@medielijah

4 жыл бұрын

lab is not your metier, try something else, no shame

@resistance110

3 жыл бұрын

@@medielijah What a stupid comment you made. Discouraging someone because they can't do a western on their first try. Stay out of labs with that negativity.

@oscarqr1

3 жыл бұрын

@@resistance110 Honestly lmao. Almost everyone messes up protocols on their first try. Hell, I still mess up protocols after a few years of lab experience. Sometimes it's funny. Sometimes I destroy weeks of work and it's not so funny. Shit happens o.0

@yeny7194

3 жыл бұрын

@resistance110 and @oscar you guys are awesome

@ronaldokevin5693

3 жыл бұрын

@@medielijah social media is not good for you, try something else, no shame

Very informative Thank you so much for taking time and explain each step.

Thank you for this video , it really helpes me to understand correctly what i'm doing, when and how. Last week was the first time i had to do this proces but it was all unclear to me. Now I know what to do . Thank you so much !

@beautifulmeena2571

5 жыл бұрын

thnku sir

@lincolnoliveira3041

3 жыл бұрын

Yuka

That final result was beautiful!

Thank You so much for your wonderful video!

all respect for this valuable work , thank you 🎩🌸

Thank you BioRad

really enjoyed watching this video, great review!!! Thank you!!!

WATCH IN X1.25 SPEED just to save you some time

@user-rf4vc7mt4d

3 жыл бұрын

Scam

@davidwaziri8460

2 жыл бұрын

Vedio

Thank you so much, it's pretty helpful

thank u so much.. this video was so helpful

when he was discarded the wash buffer or antibody, it gave me anxiety

@rowang5578

Ай бұрын

He uhh he give me butterfly 🤭 those green hand

Nice video!

Nice video about it!

Wow! This is so helpful I must say

Thanks Bio-Rad ....

@emilie6466

4 жыл бұрын

Govind Meena thanks Bio-dad...

THANK YOU.

Thanks so much! :))

Add 10 mL of primary antibody and pour off it...reminding me of the time when I sealed 2 mL of the primary antibody along with the membrane for binding and then recycled the used primary antibody...

@MrSmokingHott

5 жыл бұрын

Lamhirh amen to that!

@TheTillieTube

5 жыл бұрын

This is how we do it in real life....

@ShakespeareCafe

5 жыл бұрын

BioRad has a big budget and can afford to pour off the Expen$ive primary antibody...you can reuse it a few times before sending it down the drain

@seetheworld6656

Жыл бұрын

How to recyle it??

@NewWesternFront

11 ай бұрын

@@seetheworld6656 put it in the blue can on your house curb

"pour off the primary and secondary antibody" me watching this: o m g, i sealed and reused for many times bcs expensive😂😭

@monishajayabalan7823

4 жыл бұрын

You are not alone 😂😂

@user-we9qg3dy5n

3 жыл бұрын

Same

@jatnarivas8741

3 жыл бұрын

Have you found the results to be equally reliable either way?

@HercadosP

3 жыл бұрын

@@jatnarivas8741 not as reliable. Have had some complete failures, but good enough for repeats to verify results. When I am ready to repeat the experiment for publication I use new antibodies

Very helpful thank you

Thank you

Thanks !

Nicely explained 👏💐thnku

Thank you sir

10ml of primary Ab and 10ml of substrate volumes were SO different, this does not feel right. I've never done a western blot, can someone explain which one was wrong? It kind of looks like 100ml of substrate

What is the composition of blotting buffer, blocking buffer, wash buffer?

I have a question that might be answered here. I'm trying to get by cheaply and was wondering if I could run a western blot gel in a DGGE electrophoresis chamber ?

Thanks

Thank u soooooooooooo much🙏

it helpful video. thnx

Cool gloves!

I see this for sale on the wards scientific , but says the kit does not include fish samples, I can't find where I am suppose to buy the fish samples from?

Thanks for this video. It's been a while since I did my last WB, so thanks for a short refreshing. But don't you think the 10 mL of substrate solution were significantly more than the 10 mL of both antibody solutions? ^_^

Hi, I want to know what kinds of Substrate you used? The blue color you can see through the naked eyes or have you used a tool to analyze?

Do you use nitril gloves? Are they resistant in contact with methanol in the Buffer?

wow!!!!! great

Reminds me of developing film.

@-..l

4 жыл бұрын

John Smith I agree, like developing film. Expect, it is not in a dark room.

@JM.5387

3 жыл бұрын

In the old days, we used photographic film, and developed it in a dark room. Everything was labeled with radioactivity.

How much duration of time take to diagnose western blot test for hiv in lab???(not about window period)

good

👍 Helpful

it is 2018 and something has been updated ( instructions and so on)

Helpful video, thank you. A piece of question, though; I expected chemiluminescent detection or use of detection equipment in general to visualize the bands after the secondary antibody but the video shows direct color development through substrate addition, would you clarify?

@user-fn7hs5yc2h

7 жыл бұрын

***** Thank you for the reply, point clarified.

@diogenesofsinope2692

Жыл бұрын

@@user-fn7hs5yc2h explain pls

a rocking board is that really necessary?

why we add scondry antibodies????

could anyone tell me what are the components of the substrate he added in the final step? it can direct see the band without exposure on machine. because for me ,after adding second antibody and washing, i incubate the membrane with immunostar or Chemi lumi One L, and then exposure.

@yifanhu1546

8 жыл бұрын

+BioRadLifeScience i see, thank you so much!

@tony232cool

4 жыл бұрын

how do yo do detection after electrotransfer.

Im trying to buy one of the plastic cases that you build the transfer casette shown in the above video but cant seem to find anyone selling.. what are they actually called?

@BioRadEducation

4 жыл бұрын

Hi, Stephen! Please see www.bio-rad.com/en-us/product/mini-trans-blot-cell#fragment-4. If we can help further, call us at 1-800-4BIORAD and we'll help you find what you're looking for.

Thank you this vedio

Why did I go to grad school there are like 50 steppppSSSS

Preparation of the sample is not included?

He incubate the blot with primary antibody for 15 minutes...can we incubate for longer period like overnight at 4c

@kundansolanki33

2 жыл бұрын

For sure

hola chicos si estáis viendo esto suerte el viernes en TMB

What is the role of (HRP) in the process of western blotting?

@ashleyseal1200

7 жыл бұрын

HRP (horse radish peroxidase) is conjugated to the secondary antibody and when the membrane is incubated with a substrate, the proteins of interest can be detected. Some secondary antibodies are conjugated to a florescent molecule, in which case there is no need to incubate with substrate. Or you could just google it.

@sssaq

7 жыл бұрын

Thanks. Biochemistry is very complicated, specially if you're not taking it in your first language!

@ashleyseal1200

7 жыл бұрын

Hope I helped! Take a look at the abcam western blot video (very straight forward) or have a look on abcam.com if you're still struggling

@sssaq

7 жыл бұрын

BioRadLifeScience Thank you very much indeed.

Grt video! Can the antibodies be reused?

@puspanjali2464

Жыл бұрын

Yes you can

Why do we need two antibodies? (sorry for this dumb question, im just getting into research)

@BioRadEducation

4 жыл бұрын

Hi, Luyan! The first antibody, called a primary antibody, detects the target of interest (ex. rabbit anti-GFP). The second antibody can amplify the signal and, by more generically targeting the species that generated the antibody (ex. mouse anti-rabbit), is most cost-effective than generating reporter antibodies specific to a particular target. Visit www.bio-rad.com/classroomresources for FREE animations on how ELISA works - western blotting works because of the same target and signal amplification principles.

Why haven't you added any blocking solution?

can i use salmon sperm DNA for blocking solution?

what is the purpose of the blocking solution?

@nicheng1927

8 жыл бұрын

+S Gill preventing nonspecific binding of primary and secondary antibodies in downstream steps. Blocking agents work by covering the unoccupied areas of the membrane with a dense layer of molecules. Blocking agents can either contain proteins, or be protein-free. I use to fatty-free milk(the cheapest and common one) or chick serum to be blocking buffer. If you want to improving your sensitization in phos-antibody, there have various commercial blocking buffer for that.

One of the worse nightmares of basically every molecular biology researcher...

유투브 알고리즘 무엇...

Spr

Thenk you ,l m from in algeria l want dot blot steps

Why do we need to add substrate?

@annakrahbichler

4 жыл бұрын

the secondary Anitbody is marked, with HRP (Horseraddish Peroxidase), to gat a signal you need to add the substrate

what the purpose of the rocking platform

@antoniofernandes7816

4 жыл бұрын

I'm not sure but it might be to maintain the piece covered but with low quantity of the solution used

I am here , for my exam preparation ..

@Kingg_45

Жыл бұрын

Which exam

@AnjuVerma-mz2rx

Жыл бұрын

@@Kingg_45 3rd semester biotechnology exam...

can someone explain to me the theories of each step (why we do what we do)?

@BioRadEducation

4 жыл бұрын

Hi, Luyan! There is a lot to explain in a reply. You may want to check out our textbook "Biotechnology: A Laboratory Skills Course" available in digital and print formats for the science and theory of western blotting and other lab techniques. Visit www.bio-rad.com/textbook for more information.

Blocking solution is milk

Infibulation is necessary? Why

Most disgusting sandwich I've ever tasted.

@jatnarivas8741

3 жыл бұрын

If you tried this one, you should really try the sandwich ELISA XD

*Strange that you place the nitrocellulose in with the gel while electrophoresis occurs. I'm not familiar with the technique... but:* 1) If the proteins can electrophoretically move along the nitrocellulose, why bother with the gel at all? 2) If the proteins can't electrophoretically move along the nitrocellulose, then surely they must blot in to it and stop migrating throughout their elecrophoretic journey, which would produce streaks of misplaced proteins.

@Jamieishere1

9 жыл бұрын

***** Thanks for the clarification. I must have not watched the video thoroughly enough. My mistake.

@tony232cool

4 жыл бұрын

electrophoresis with gel only and then electrotransfer with gel ad membrane.

EL RODILLITO ME CAUSO GRACIA sjjsjs

Meeting Dr Igudia KZread channel was the beginning of a new life for me after using his herbs medication in curing my Genital herpes Virus

An Excel file with the sequential steps and links to the video timepoints should be added as downloadable link

X

8:10 Yeah, how about we don't pour $200 down the drain...

@mitylene_bailey

5 жыл бұрын

What should you do with it?

@jatnarivas8741

3 жыл бұрын

@@mitylene_bailey I read comments of people sealing it and reusing it at another time.

is it so much to ask that you put a goddamn + and - on the anode and cathode side, instead of saying "color coding to ensure proper orientation" in the manual but never actually explain what the color coding is?

@marios1861

3 жыл бұрын

the color coding is pretty uniform. red for anode and black for cathode. sometimes black can be white or yellow or green but red stays pretty much the same.

や11

You must love Jehovah your God with all your heart and with all your soul and with all your mind and with all your strength. You must love your neighbor as yourself. Jesus the anointed is Lord! Repent and be baptized and believe the Gospel. Work out your own salvation with fear and trembling.

@Verspassungsschutz

4 жыл бұрын

Shut Up

Thanks!!!