Thanks for the suggestion Arne.  Have you used this Ab for marking plasma membranes for segmentation before?  What has been your experience with this reagent?

Much appreciate the feedback.

Best,

--
Samuel A. Connell
Director of Light Microscopy
Cell & Tissue Imaging Center
St. Jude Children's Research Hospital
262 Danny Thomas Place
Memphis, TN 38105-3678
Office (901) 595-2536
Cell (901) 603-3162
[log in to unmask]




On 8/17/09 10:43 AM, "Arne K Christensen" <[log in to unmask]> wrote:

The Developmental Studies Hybridoma Bank has a nice monoclonal antibody
against the alpha subunit of the NKAtpase.  It's called alpha5 (a5).

http://dshb.biology.uiowa.edu/a5




Connell, Samuel wrote:
> Tim,
>
> Do you have a particular Ab to recommend for the sodium-potassium ATPase you mentioned?
>
> Thanks,
>
> --
> Samuel A. Connell
> Director of Light Microscopy
> Cell & Tissue Imaging Center
> St. Jude Children's Research Hospital
> 262 Danny Thomas Place
> Memphis, TN 38105-3678
> Office (901) 595-2536
> Cell (901) 603-3162
> [log in to unmask]
> ________________________________________
> From: Confocal Microscopy List [[log in to unmask]] On Behalf Of Timothy Feinstein [[log in to unmask]]
> Sent: Thursday, August 06, 2009 8:41 PM
> To: [log in to unmask]
> Subject: Re: Whole Cell Dye.    .
>
> Hi Cam,
>
> In the past we have had good results with chloromethylfluorescein
> diacetate (CMFDA).  CMFDA is an intravital dye that fluoresces in the same
> channel as GFP, however as far as I know only works in live cells.  If
> transfection is possible, people often use GFP-family proteins as a
> co-transfection marker.   These pass freely through nuclear pores and
> therefore produce even cellular labeling.  If an antibody is preferable,
> the sodium-potassium ATPase marks the plasma membrane as clearly and
> evenly as anything that I have seen.
>
> All the best,
>
>
> Tim
>
> Timothy Feinstein, PhD
> Postdoctoral Associate, Vilardaga laboratory
> University of Pittsburgh Dept. of Pharmacology and Chemical Biology
> Pittsburgh, PA
>
>
>
>> Hi List
>>
>> Can anyone recommend a dye (or antibody) that will stain the whole cell
>> evenly to the plasma membrane? I need a dye that will give fairly even
>> staining across the whole cell to make segmentation easier.
>>
>> I have tried CellMask but is it a bit variable in its staining pattern,
>> there are blobs in the cytoplasm and nucleus may or may not stain up.
>> Something that stains just the plasma membrane would also be suitible. I
>> have used the invitrogen FX dyes before (not for this experiemnt) and
>> have found them a bit hit and miss too. I have also tried WGA but it
>> doesn't give an even stain around the mebrane which makes segmentation
>> quite dificult.
>>
>> Any suggestions would be greatly appreciated.
>>
>>
>> Thanks
>>
>> Cam
>>
>>
>> Cameron J. Nowell
>> Microscopy Manager
>> Central Resource for Advanced Microscopy
>> Ludwig Institute for Cancer Research
>> PO Box 2008
>> Royal Melbourne Hospital
>> Victoria, 3050
>> AUSTRALIA
>> Office: +61 3 9341 3155
>> Mobile: +61422882700
>> Fax: +61 3 9341 3104
>> Facility Website
>>
>>
>>
>>
>> This communication is intended only for the named recipient and may
>> contain information that is confidential, legally privileged or subject to
>> copyright; the Ludwig Institute for Cancer Research Ltd does not waiver
>> any rights if you have received this communication in error.
>> The views expressed in this communication are those of the sender and do
>> not necessarily reflect the views of the Ludwig Institute for Cancer
>> Research Ltd.
>>
>>
>>
>
>
> Email Disclaimer:  www.stjude.org/emaildisclaimer
>
>


--

__

Arne K Christensen

Postdoctoral Research Associate
University of Massachusetts, Amherst

USGS Conte Anadromous Fish Research Center
One Migratory Way, PO Box 796
Turners Falls, MA  01376

Email: [log in to unmask]
Phone: (413) 863-3827
  Fax: (413) 863-9810
  URL: www.biobog.com