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| Cell Biology and Cell Culture 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
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| 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
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| 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
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#4
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| 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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| fiber , mitotic , spindle , stain |
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