Phospho -serine STAT specific antibodies.

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samm
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Phospho -serine STAT specific antibodies.

Is anyone aware of/has used any phospho-SERINE STAT (eg Stat6) specific antibodies. I'm NOT looking for the phospho-tyrosine antibodies. Phosphorylation of serine residues actually negatively regulates function, while on tyrosine residues, it activates!

Richard Taylor
Richard Taylor's picture
Phospho-serine STAT6

Phospho-serine STAT6 Antibodies are not yet available from any major suppliers, however phosphoserine Abs for the other STATs are available:

http://www.abcam.com/index.html?datasheet=4742
http://www.abcam.com/index.html?datasheet=4747

I don't know what you're trying to do but you could be inventive with your use of generic phosphoserine Abs.

samm
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Thanks! I was planning to try

Thanks! I was planning to try out some - we suspect both p-Tyr and p-Ser involvement in the pathway we are working on, so doing an IP with Stat6, followed by a probe with generic phospho serine Abs is definitely worth a shot.

nin1318
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i have used a phospho-ser.

i have used a phospho-ser. stat3 antibody before...i don't remember right now who it was made by, but i'll look into it and post back.

samm
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I'd really appreciate that -

I'd really appreciate that - could not find any direct refs. Am most interested in Stat6, Stat3 and Stat5, in that order.

TRMADR
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samm wrote:I'd really

samm wrote:

I'd really appreciate that - could not find any direct refs. Am most interested in Stat6, Stat3 and Stat5, in that order.

Hi Samm, the concept of using some form of pan-specific capture antibody for a STAT, (or against whatever specificity you are looking at) followed by a detect of a generic phospho antibody (as Richard mentioned) is something that we have tried once before. In theory it should work, although there may be some steric hindrance, or other issues that cause the assay to fail. I would be interested in finding out if this works or not, as many companies go to great lengths to create phospho-specific ELISA assays and detection systems. If all else fails, have you considered a custom polyclonal antibody? Many companies offer them--and are quite competitive. If you were to consider this, i would suggest only that you ensure that the company produces their own antibodies. These days a lot of companies are just reselling the services of others--not that this is necessarily bad, but for cost containment, and the best service definately chose a company that keeps that service inhouse.
-d

samm
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I'm planning to try it out

I'm planning to try it out using the Santa Cruz SC20 rabbit polyclonal against Stat6, followed by a phospho-serine Ab raised by another lab here. (Have also used the mouse monoclonal pY-Stat6 Ab from these guys - it works very well)
Could you let me know of any of the custom Abs companies that you have tried out.

samm
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Also, would much rather

Also, would much rather prefer Abs that work in Westerns, rather than solution phase (ELISA/FACS etc) for this set.

TRMADR
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samm wrote:I'm planning to

samm wrote:

I'm planning to try it out using the Santa Cruz SC20 rabbit polyclonal against Stat6, followed by a phospho-serine Ab raised by another lab here. (Have also used the mouse monoclonal pY-Stat6 Ab from these guys - it works very well)
Could you let me know of any of the custom Abs companies that you have tried out.

Hi samm, in the interests of fairness, I also work for an antibody company. The concept that Richard proposed is something that i tried once, using one of our products and the well known phos-tyr antibody (I forget the clone name, but Upstate carries it). The time i tried it, i didnt get any results, but in all fairness it was done using a limited amount of ab, and was not repeated. On the topic of custom solutions, you are welcome to contact me offline on matter (just click my name and send me an email). I can provide you with whatever information you would like. we do this often. Another advantage is that some companies work on a collaborative basis, which may further reduce your costs--i.e. they make it and you test it, or you provide the antigen and they make it.

samm
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Thanks! I'll try this expt,

Thanks! I'll try this expt, as well as genetic mutants, and see what happens. I'm moving down to UCS in 2 months, so will start custom Abs after that. Would prefer confirmation by independent methods. Have you guys ever tried making specific phospho-serine Abs - is it hard to do?

nin1318
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santa cruz has many stat

santa cruz has many stat antibodies...these are the ones that i used previously, and had success with them.

http://www2.scbt.com/catalog/action.lasso?-response=product_search_list.html&-Error=catalog%2Fno_results.html&-token.order_id=4202867&search_field=stat3&-nothing.x=0&-nothing.y=0&-nothing=go

sorry the link is so long!

nin1318
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i guess it didn't matter..i

i guess it didn't matter..i was worried that the link being on multiple lines would make it cut off part when it made it active.

samm
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Thanks! Sadly those are the

Thanks! Sadly those are the regular STAT or pY STAT Abs - did not have muck luck with a direct supplier, or usig "general" phosphoserine Abs. Am looking out for alternative ways of experimentation - probably reporter assays.

nin1318
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looks like there are several

looks like there are several phospho-serine abs there:

p-Stat3 (Ser 727)-R: sc-8001-R
p-Stat4 (Ser 721) sc-16317
p-Stat4 (Ser 721) sc-22160

sorry if these are of no use.

nin1318
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there are also stat3 serine

there are also stat3 serine abs from calbiochem and cell signaling...probably others as well. i havent seen any for stat 5 or 6 though.

samm
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Thanks! The Stat3 one might

Thanks! The Stat3 one might be of use, though I'm right now focused primarily on 6, (3&5 are next!). Have you used this Ab?

nin1318
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i used one from santa cruz..

i used one from santa cruz...i haven't tried the ones from cst or calbiochem. had good luck with it. unlike the phospho-tyr which was not easy to get.

suzanoberle
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samm wrote:Is anyone aware of

samm wrote:

Is anyone aware of/has used any phospho-SERINE STAT (eg Stat6) specific antibodies. I'm NOT looking for the phospho-tyrosine antibodies. Phosphorylation of serine residues actually negatively regulates function, while on tyrosine residues, it activates!

I work for a company called MBL and we have a phospho-Ser727 STAT3 antibody. We also have phospho-STAT6 but it's Tyr 641, not Ser, so it isn't quite what you're seeking.

No sales pitch intended, but if you're interested, go to www.mblintl.com and type jm-3474 in the search engine.

samm
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Thanks! Have already got the

Thanks! Have already got the Stat3 ones, will try out the MBl ones too. Is there any hope for the serine Stat6 Abs? Using a "general" p-Serine Ab does not work!