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Mitotic spindle fiber stain

Mitotic spindle fiber stain - Cell Biology and Cell Culture

Mitotic spindle fiber stain - Cell Biology Forum. Cell Culture Forum. Post and ask questions about cell culturing, cell lysis, cell transfection, cell growth, and cell biology.


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  #1  
Old 04-07-2006, 09:27 PM
Vincenzo Petix
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Default Mitotic spindle fiber stain



Hi,
I'm a biologist who works in a genetics laboratory.
I'd like know if exists a method for stain mitotic spindle fibers
without use rabbits antibody
someone can help me? Thanks





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  #2  
Old 04-09-2006, 08:46 AM
Sephy
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Default Mitotic spindle fiber stain

Just search for the specific probe from Molecular Probes, devoured by
Invitrogen.

However, it would cost you about 500+ dollars...

To stain mitotic spindle fibers perhaps you'd better find a
tubulin-specifice probe.

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  #3  
Old 04-09-2006, 10:23 AM
Vincenzo Petix
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Default Mitotic spindle fiber stain

you didn't solve my problem ,but thank you very much for giving me your help

"Sephy" <[Only registered users see links. ]> ha scritto nel messaggio
news:1144572370.535129.102070@g10g2000cwb.googlegr oups.com...


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  #4  
Old 04-10-2006, 08:37 AM
Sephy
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Default Mitotic spindle fiber stain

Uh...Well, maybe there are commonly 3 ways to stain microtubules as
well as other components of cytoskeleton like F-actins. It is the
purpose of you experiment that decides.

1.Stains with labelled probe,usually fluorescence labelled.It is
readily obtain from many vendors like Invitrogen or Sigma.In most
occasions,you can get a very nice photo from that.However it will cost
you much,and the standard protocols are not directly applicable-I
suffered from this recently so believe me.And the main drawback is you
can hardly stain a cell while it's living.So this method is eligable in
morpholgical research but unavailable when you want to do vital stain.

2.Immunofluorescence.Average cost,average effort required and quite
nice photos obtainable(by quite nice I mean it surely do some harm to
cells).There are tons of protocols about this method.I feel strange why
you aviod it.But it is you make decision.

3.GFP,recently GFP-mTn.As to label mitotic spindles,which mainly
contain microtubules,you could construct a fusion protein with
alpha-tubulin and GFP/GFP-mTn,express it in vivo,excite it,say cheese!
It's a perfect method because least harm,if not none,is done to
cells.Not only that,also there's no need trying to persuade your boss
to pay the bill for the kit,and you don't have to kill lovely bunnies
only to prepare antibodies.There are many articles on PubMed to which
you could refer.

Hope it would be useful.

I am now involved in a work on actin cytoskeletonin plant.Unfortuately
I am experiencing protocol frustrations.Fluorescence phalloidin from
Invitrogen sucks.The protocol enclosed is somewhat a piece of bullshit.

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