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Our Abpromise guarantee covers the use of ab14692 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|WB||1/200 - 1/1000. Detects a band of approximately 42 kDa (predicted molecular weight: 38 kDa).|
|IHC-FoFr||1/500. PubMed: 24846136|
|IHC-Fr||Use at an assay dependent concentration.|
|IHC-P||1/1000. PubMed: 19096824|
Immunohistochemical analysis of formaldehyde-fixed, paraffin-embedded human CNS sections labelling Synaptophysin with ab14692 at a dilution of 1/1000. The secondary antibody used was biotin conjugated polyclonal goat vector at 1/250. Antigen retrieval was heat mediated using citrate.
ab14692 staining Synaptophyson in mouse retina (8 weeks old) and brain tissue sections by Immunohistochemistry (IHC-Fr - frozen sections). 10µm frozen sections were fixed with 4% paraformaldehyde for 10 minutes and blocked with 10% donkey serum for 20 minutes at room temperature. Samples were incubated with primary antibody (1/100 in PBS) overnight at 4°C. An Alexa Fluor 488®-conjugated donkey anti-rabbit IgG polyclonal (1/1000 in PBS) was used as the secondary antibody (1 hour at room temperature). Counterstained with Hoechst 33258 (blue) for 10 minutes at room temperature.
Images shows: GCL - ganglion cell layer, IPL - inner plexiform layer, INL - inner nuclear layer, OPL - outer plexiform layer, ONL - outer nuclear layer; DG - dentate gyrus region of the hippocampus.
Synaptophysin antibody (ab14692) western blot on mouse forebrain extract (10µg protein/lane). The blot was exposed for 10 minutes to autoradiographic film. Ladder markers from 7-194 kDa.
Lane 1 = 1/500 ab14692
Lane 2 = 1/200 ab14692
ab14692 staining cultured ED18 rat primary hippocampal neurons by ICC/IF. The cultured cells were fixed with 4% paraformaldehyde for 5 minutes and blocked with 10% donkey serum in 0.1% PBS-0.3% TritonX for 30 minutes at 24°C. The cultured cells were then stained with ab14692 at 1/200 in 0.3% TritonX with 0.1% PBS and 10% donkey serum for overnight at 4°C. An Alexa Fluro 488 donkey anti-rabbit polyclonal antibody at 1/1000 was used as the secondary antibody. Nuclei were stained with 1.43µM Hoechst and can be observed in blue. Synaptophysin expressed in axon, dendrites and particularly carrying in the vesicles.
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