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@  clutz : (18 December - 04:36 AM) Hi everyone,
In June 2012 I bought a number of antibodies from AbD Sertotec and, according to the data sheet from the STAR9B conjugate dated June 2012, the recommended working dilution should be between 1:200 and 1:500. but, in a later datasheet (from november 2013) the recommended working dilution is 1:25-1:100.
My student asked the Serotec technical support and they told her to use the later STAR9B dilutions (2013) for our flow cytometry experiments: Bovine PBMC surface stained with purified Mo anti-Bov CD4+ Rabbit anti-mo IgG F(ab´)2-FITC and an intracellular staining with mo anti-bo IFNg-PE.
We have got signals from IFNg in every sample and controls. We ran a test with a mo anti-bo CD4 FITC + mo anti-bo IFNgPE; and another test blocking with normal mo serum after de STAR9B staining and the stain with mo anti-bo IFNgPE, and the effect dissapeared, suggesting that the STAR9B is capturing the second conjugate as well... Is that possible??? Is it a matter of wrong dilution of STA9B conjugate and monovalent binding of STAR9B to de primary antibody?
Please, help me to solve this mystery....
Thanks in advance
@  pito : (16 December - 01:19 PM) Hallo.
@  student47 : (16 December - 07:27 AM) hi
@  Trof : (06 December - 07:51 AM) Nothing tells you more "have a nice weekend" like a spill of ml of beta-mercaptoethanol on the bench by a new colleague.
I'm going to kill him as soon as my headache ends..
@  vetticus3 : (29 November - 01:53 AM) different epitopes... all poly... all produce really beautiful IF... picking one and hoping for the best. :D
@  mdfenko : (28 November - 12:54 PM) they may be for different epitopes, poly and monoclonal, native and/or denatured protein (western blot or elisa/ip compatible
@  vetticus3 : (28 November - 03:19 AM) Anyone have a good idea on how to pick a good antibody? The same comp has 3 for the same protein - getting confused. :S
@  jerryshelly1 : (27 November - 02:33 PM) Everyone have a good Turkey Day!!
@  Inbox : (21 November - 12:11 PM) @Trof: nice. finishing 15th hrs non-desk work here.
@  Trof : (21 November - 12:47 AM) I just spend six hours writing. Need tea. And yes, it's 10 in the morning, here.
@  gfischer : (20 November - 07:27 AM) @Curtis And remember that a little stroking of the ego can never hurt
@  pcrman : (19 November - 06:30 PM) @Curtis just tell the other party what you intend to do, what you can offer, trying to get one interested and willing to work with you.
@  Curtis : (18 November - 09:31 PM) want to write a collaboration request. Not sure how to write it.
@  Curtis : (18 November - 08:08 AM) I never liked plate-based PCR machines.
@  Trof : (15 November - 02:16 PM) Whole new work experience.. unsticking optical PCR plate melted to the heated cover..
@  Curtis : (11 November - 12:32 AM) I'll make sure my zombies don't eat czech chiks
@  Trof : (10 November - 01:01 PM) @Curtis And the humankind is doomed! ;)
@  Curtis : (06 November - 06:04 PM) Ph
@  autophagator : (05 November - 11:48 AM) fenotipic supression
@  Curtis : (03 November - 01:06 AM) the design was difficult

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