inter Spem et Metum

I kocked into this post the other day while I'm skimming over my Google Groups.I thought it was somewhat relative to my recent work so I replied in the thread.The next day I was informed of his reply indicated that I didn't hit. So I organized as much as I know to compose a second reply.And this seemde to satisfy him.

After re-read what I wrote,I was convinced that for sure there're still tons of flaws in my knowledge and comprehension although I didn't convince myself of what and where the flaws are.Anyhow I must know it if I want to do something in future.After all,illusion of knowledge is more dangerous than igorance of knowledge,as Barry Marshall,the 2005 Nobel laureate in Physiology or Medicine who visited our school recently,told in the lecture.

Well,I try to wrote that reply,in case that(dreaming that) I become a big wig and be asked to wrote a review or something.

Em...Realistically,consider it a review paper for a Master's diploma...


Here I post the thread.Sephy is me.


Mitotic spindle fiber stain
All 4 messages in topic - view as tree
From: Vincenzo Petix - view profile
Date: Sat, Apr 8 2006 5:27 am
Email: "Vincenzo Petix" <v.pe...@libero.it>
Groups: bionet.cellbiol
Not yet rated
Rating:
show options



Reply | Reply to Author | Forward | Print | Individual Message | Show original | Report Abuse | Find messages by this author


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






From: Sephy - view profile
Date: Sun, Apr 9 2006 4:46 pm
Email: "Sephy" <phoenix...@gmail.com>
Groups: bionet.cellbiol
Not yet rated
Rating:
show options



Reply | Reply to Author | Forward | Print | Individual Message | Show original | Remove | Report Abuse | Find messages by this author


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.








From: Vincenzo Petix - view profile
Date: Sun, Apr 9 2006 6:23 pm
Email: "Vincenzo Petix" <v.pe...@libero.it>
Groups: bionet.cellbiol
Not yet rated
Rating:
show options



Reply | Reply to Author | Forward | Print | Individual Message | Show original | Report Abuse | Find messages by this author


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

"Sephy" <phoenix...@gmail.com> ha scritto nel messaggio
news:1144572370.535129.102070@g10g2000cwb.googlegroups.com...



- Hide quoted text -
- Show quoted text -

> 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.









From: Sephy - view profile
Date: Mon, Apr 10 2006 4:37 pm
Email: "Sephy" <phoenix...@gmail.com>
Groups: bionet.cellbiol
Not yet rated
Rating:
show options



Reply | Reply to Author | Forward | Print | Individual Message | Show original | Remove | Report Abuse | Find messages by this author


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.