Re: [Histonet] IF staining on peritoneal macrophages

else, Andrea     --- On Wed, 3/3/10, Mauricio Avigdor bitesizell...@gmail.com wrote: From: Mauricio Avigdor bitesizell...@gmail.com Subject: Re: [Histonet] IF staining on peritoneal macrophages To: Andrea T. Hooper andreahoo...@rocketmail.com, histonet@lists.utsouthwestern.edu Date: Wednesday

Re: [Histonet] IF staining on peritoneal macrophages

block.   If you want to share your protocol, maybe I can give you some further tips?   Let me know if I can help further, Andrea   --- On Wed, 3/3/10, Mauricio Avigdor bitesizell...@gmail.com wrote: From: Mauricio Avigdor bitesizell...@gmail.com Subject: Re: [Histonet] IF staining on peritoneal

Re: [Histonet] IF staining on peritoneal macrophages

Rat anti-mouse F480 from serotec works hwell. It is crucial that you use a secondary against rat IgG that is highly cross adsorbed to many species, including mouse. I suggest whole IgG of donkey anti-rat IgG from Jackson Immunoresearch. Just make sure to pick the version adsorbed against mouse.

Re: [Histonet] IF staining on peritoneal macrophages

Rat anti-mouse F480 from serotec works hwell. It is crucial that you use a secondary against rat IgG that is highly cross adsorbed to many species, including mouse. I suggest whole IgG of donkey anti-rat IgG from Jackson Immunoresearch. Just make sure to pick the version adsorbed against mouse.

Re: [Histonet] IF staining on peritoneal macrophages

Hi Andrea, Jackson lists a couple of FITC-conjugated donkey anti-rat secondaries: 712-095-150http://www.jacksonimmuno.com/MERCHANT2/merchant.mv?Screen=PRODProduct_Code=712-095-150 - Whole Donkey Anti-Rat IgG (H+L).

[Histonet] IF staining on peritoneal macrophages

peritoneal macrophages? ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet

Re: [Histonet] IF staining on peritoneal macrophages

...@gmail.com wrote: From: Mauricio Avigdor bitesizell...@gmail.com Subject: [Histonet] IF staining on peritoneal macrophages To: histonet@lists.utsouthwestern.edu Date: Thursday, February 18, 2010, 11:00 PM Greetings all, I am trying to do immunofluorescence on peritoneal macrophages. I am having

[Histonet] IF staining on peritoneal macrophages

Thank you Adam and Jay for your replies. Cytoslides are the pre-marked slides for use with the Cytospin - http://www.thermo.com/com/cda/product/detail/0,1055,21035,00.html Adam, do you have a recommended concentration of BSA for this? Also, do you air dry the slides prior to fixation? Do you air

Re: [Histonet] IF staining on peritoneal macrophages

When I did it, I think I used our standard sort buffer, which is 0.2% BSA in PBS pH 7.4. I've also read that you can use anything with serum, typically 5-10%. The key seems to be prewetting the membranes with something with protein. I usually cytospin them and briefly (30 seconds) air dry them

[Histonet] IF staining on peritoneal macrophages

You may want to try using the NuView Cyto prep technique from QC sciences it is a good alternative to a Cytospin ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet

[Histonet] IF staining on peritoneal macrophages

Greetings all, I am trying to do immunofluorescence on peritoneal macrophages. I am having a couple of issues that I was hoping one of you could help me resolve. Firstly, I am having uneven results with the Cytospin. Cells tend to get washed off the slides during rinses. Does anyone have tips on

Re: [Histonet] IF staining on peritoneal macrophages

The best way I know to get cells to stick is to first cytospin liquid with some protein (BSA or serum) onto the filters to pre-wet them. After that's done, cytospin your cells. I've gotten this to work with fairly rare FACS sorted cells. I've also found that the smaller volume the cells are in,

Re: [Histonet] IF staining on peritoneal macrophages

Mauricio, Sorry if this is too obvious, but are you using plus slides? I'm not sure what you mean by cytoslides. I always use plus slides with the Cytospin, and they stay on through HE, Pap, and DQ, although I'm not trying IF on them. Jay A.