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I think that my purchasing agent was not clear. The antibody has not worked for me yet, but my current complaint is that Federal Express did not handle the shipping properly. It was shipped by you Fed Ex overnight but sat at Fed Ex for an additional day. While I presume that the Ab should be stable, I have no idea how it sat at Fed Ex. In the meantime: Description of the problem: (no signal, high background, non-specific signal etc). No signal Sample (Species/Cell extract/Nuclear extract/Purified protein/ Recombinant protein etc): Fixed MDCK cells Sample preparation: Fixed with 4% paraformaldehyde, permeabilized with 0.5% Triton X100, blocked with fish skin gelatin Blocking Conditions (Buffer/Time period, Blocking agent etc) See above Primary Antibody (Manufacturer/Species/Diluent/Dilution/ Incubation time, Wash step) YOUR Ab, at recommended 1:250 dilution in PBS- with fish skin gelatin and saponin Secondary Antibody (Manufacturer/Species/Diluent/Dilution/ Incubation time, Wash step) Molecular Probes, goat anti chicken 546, 1:100 Detection method Immunofluorescence Positive and negative controls used. (Please specify) Positive control: same cells, with expression induced under the same conditions, good signal with 9E10. Negative control: same cells, expression repressed Optimization attempts (problem solving) Have you run a No primary control? I usually do this when I have too much background, not no signal How many times have you tried this assay? Once-I will try it again but wanted to raise the concern re Ab sitting at Fed Ex.
Asked on Mar 08 2004
I looked at the tracking for Fed Ex and it states that the package was delivered on Feb 25 at 11am and signed for by L. Kim. Although the antibody arrived a day late, it should not affect the antibody (it should have still been cold). If you continue to experience problems with this antibody, please contact me again.
Answered on Mar 09 2004