Immunofluorescence

Рет қаралды 271,612

14 жыл бұрын

( www.abnova.com ) - Immunofluorescence is a technique to visualize a specific protein or antigen in cells or tissue sections by binding a specific antibody chemically conjugated with a fluorescent dye. We use the indirect immunofluorescence staining to perform cells fixed on slides and examine under a fluorescence microscope. More videos at Abnova www.abnova.com

Пікірлер: 45

@missroua6862 8 жыл бұрын

simple and clear, thanx a lot

@roaahashim9373 8 жыл бұрын

amazing ! lots of thanks

@BaughbeSauce 10 ай бұрын

wow. so many steps!

@noratiqahjusril6981 5 жыл бұрын

how to deal with cells culture at different concentrations using this method?

@redSHIFT69 13 жыл бұрын

thanks abnova

@wiraniaritiasnitha9419 3 жыл бұрын

Thank you

@alexandreluizborgessilva6018 3 ай бұрын

to fix the antigen, can I replace the paraformaldehyde with formaldehyde?

@danmiller2177 3 жыл бұрын

Thanks!!!

@bish4008 7 жыл бұрын

Thanks

@moshiif 14 жыл бұрын

Thanks a lot :)

@saivikra 11 жыл бұрын

thanks

@user-qh5ch3wy3e 3 жыл бұрын

Could u please tell me what is the membrane applied for antibody incubation?

@ericwang1245 Жыл бұрын

parafilm

@hlainghlaingmyint645 10 жыл бұрын

Thanks! Tokyo Institute of Technology

@rehabhamdy3341 11 жыл бұрын

thanksssssssss a lot

@boyinalabcoatboyinalabcoat393 2 жыл бұрын

Why to use blocking solution before the primary antibodies? Is there not a chance you will be blocking the protein you want to find? Or it is just worth it, because even if you block some of the target proteins, the primary antibody will just do better in finding the not blocked target proteins, than finding inespecific conections?

@TahseenRaza_KT1983 2 жыл бұрын

If you do Blocking properly you will get good result with nonspecific background. so better do blocking and you will get the clean result and you do not have to worry about unspecific binding that may give you false positive result. hope you will get it. Thanks

@strivingforexcellency Жыл бұрын

To make epitope more visibility

@RohithBasu 11 жыл бұрын

kool !!

@HKheir 13 жыл бұрын

this is very helpfull,thank u soo much :)

@aaryashkushwaha9450 3 жыл бұрын

Hiii

@catalinarubio8850 7 жыл бұрын

thank u ! why do u use parafin?

@andreas.chrysostomou 7 жыл бұрын

Catalina Rubio PFA is used to fix the cells and its inner structures so you can target with antibodies and be able to see them under the lens

@sazsalamat 21 күн бұрын

every nice even after 14 years

@flowehghr108 11 жыл бұрын

Thanks! What is the white thing you put the cover slip against at 1:20?

@yfn9408 4 жыл бұрын

parafilm

@rajinashakya1800 4 жыл бұрын

@@yfn9408 m confused, if parafilm is placed on top of the cells ...then how primary antibody will interact with the cells crossing parafilm??

@neuroudec 12 жыл бұрын

mmmm I'm gonna follow this exact protocol right now

@jujetaly322 7 ай бұрын

Wats the pink solution that is removed at the beginning of the video?

@tenzinglobsang8587 5 ай бұрын

The culture media most probably

@SoulBladeM 9 жыл бұрын

weird, this is nothing like the stuff in my micrioblogy textbook.

@TwiSTeDBeAnS 6 жыл бұрын

Probably because this is a mounting procedure for immunostaining eukaryotic cells. Prokaryotes don't have any organelles or other large structures inside the cell to fluorescently label and look at under a microscope so there is no need to go through with a permealization procedure like this. Even if you wanted to label something in the cell membrane/wall, bacteria are small enough that it would be very hard to distinguish puncta with the resolution of traditional microscopes, so you wouldn't really get much useful information by immunostaining. In a microbiology setting, it is typically much more useful, cheaper, and easier to place cells directly from culture to a slide, fix them to the slide with a bunsen burner, and use common stains (carbol fuschion, crystal violet, malachite green) to see bacteria and any potential structures.

@shagunsharma2812 3 жыл бұрын

Wooooow ♥️

@TJ__23 4 жыл бұрын

th u :)

@jakku71 5 жыл бұрын

Just before mounting, do the cover slips have to be dried? Or can we mount the coverslips wet?

@Daniel-wb6yl Жыл бұрын

do you have the answer already now bro? I’m also having the same question :(

@dr.tranngocthien 10 ай бұрын

same question

@Daniel-wb6yl 9 ай бұрын

It's best to leave it wet. maybe if there's a lot of water (maybe, a drop?) on the opposite side of the cell side, you can tap that side lightly to a kimwipes so the droplet won't affect your mounting. If we were to leave it too dry, the cells can be destroyed, sheared or shrunk. hope it helps. @@dr.tranngocthien

@shash1810 5 жыл бұрын

what do u mean by primary and secondary antibody

@iNailHD 5 жыл бұрын

A first antibody is needed in order to recognize the protein you want to observe. Then, you use a specific second antibody which is gonna recognize the first antibody. In the second antibody, there is a fluorochrome ( it is fluorescent in the microscope ). I think i didn't say something wrong, but i'm not sure, also sorry if my english is quite bad :)