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Our Abpromise guarantee covers the use of ab4066 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|ICC/IF||Use at an assay dependent concentration. PubMed: 23624931|
|IHC-P||1/10 - 1/50. Antigen retrieval is not essential but may optimise staining.|
|IHC-Fr||1/200 - 1/500.|
|WB||Use at an assay dependent concentration.|
ab4066 at a dilution of 1/200, staining S100 in astrocytes (Alexa 488 secondary at 1/2000) on 30
NB: No labeling observed following omission of primary antibody.
Sections were viewed using an Axioplan 2 Imaging microscope (Imaging Associates) fitted with 10x, 20x and 40x Plan-Neofluorobjectives (Zeiss, Germany) and images were taken using a AxioCam Hrm digital camera (Zeiss, Germany) and AxioVision software (Imaging Associates).
This image is courtesy of an Abreview submitted by Ms Nancy Nutile-McMenemyThe extra band at 18kDa may be a result of the protein's thermolabile properties. The band at 21 kDa maybe a dimer of s100a and s100b and the 10/15 kDa may be the subunits. However, there are also references to suggest these may be immunoreactive proteins with differing molecular weights depending on proliferative or differentiated state of the cell (Anat Embryol (Berl). 1997 Nov;196(5):403-16).
ab4066 at a 1/100 dilution staining rat cortical preps grown 14 days in culture, plated onto coverslips. The preps were acid/alcohol fixed for 15 minutes, blocked and then incubated with the antibody for 1 hour. Bound antibody was detected using a Alexa fluor ® 555 conjugated goat polyclonal antibody. Nuclei were visualised using DAPI (blue). Note: s100 is colocalizing around the nucleus.
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