PC12 differentiation NFG - (Nov/27/2006 )

Hi all,

where can i buy NFG from best recommended source?

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What is NFG?

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oops, should be NGF nerve growth factor

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You should note that PC12 cells are different from lab to lab - the PC12 cells in our lab for example do not differentiate in response to NGF. Good luck...

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we buy NGF at www.axxora.com

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I used NGF to treat my PC12 it worked very well. After 2-3 days you see very long and beautiful processes when stained the cell afterward.

I bought the NGF from Harlan Bioproduct, I use the (2.5S NGF)

I used it at a concentration of 50microgram per ml

It comes in a powder form, you have to reconstitute it in 100mM Na Acetate pH 4.5


Good luck
J

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Hi jmo,

what stain did u use for the dendrites ?

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i guess MAP-2 is for dendrites and tau is for axons. we use 100ng/ul, some PC are not easy to differenciate as said before. dont forget to decrease the FBS and horse serum concentrations.

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Hi. We use MAP2 too, it works pretty good, but only at high dilutions. If im not mistaken though, the PC12 cells do not grow real dendrites or axons, but neurite-like extensions containing neuronal specific tubulin (MAP2 binds to that).

Anyways, I have a question myself. How do you fixate the PC12 cells? We tried collagen coated coverslips, but there were still very few cells attached to it. Now we fixated the cells directly in our (collagen coated) petri-dishes with 4% formaldehyde; cell yield is great. However, staining the cells is a pain. We first had the idea to cut/melt the dishes into microscope-slides-sized-squares using a hot scalpel. While this works for say 2 or 3 dishes, now we have 32 Do you think it's possible to directly treat the dishes with antibodies by marking areas with a dakocytomation pen eg. ? Or any other ideas?

Thanks in advance.

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I have used glass bottom dishes in the past (http://www.glass-bottom-dishes.com). Basically they're dishes or plates with holes cut in the bottoms. The holes are then covered from the underside with glass coverslips. You can get them coated or uncoated. You can then culture your cells almost exactly as you normally would, fix, and process for immunoreactivity in the well. After immunoloabeling you can dissolve the glue holding the glass coverslip in place and mount it on a slide for viewing and long term storage.

You can always try the free sample to evaluate for your studies. PM me if you have any questions and good luck.

ps. I will be working with PC12s very soon and was wondering what you found to be the best matrix for adhesion.

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