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Human monocytes and U397 cell line.
Our Abpromise guarantee covers the use of ab205 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
ab91545 - Mouse monoclonal IgM, is suitable for use as an isotype control with this antibody.
|IHC-R||Use at an assay dependent concentration.|
|WB||1/100 - 1/500. Detects a band of approximately 43 kDa. As this is a mouse IgM antibody, an anti-mouse IgM secondary antibody must be used to detect this primary.|
|IHC-Fr||Use at an assay dependent concentration.|
|IP||Use at an assay dependent concentration. PubMed: 27336173|
Immunocytochemistry/ Immunofluorescence analysis of ARPE cells labelling F-actin with ab205. Cells were fixed in 4% paraformaldehyde for 10 min, washed with PBS, and blocked for one hour. Cells were then incubated for 3h at 37°C with Anti-F-actin antibody [NH3] (ab205) at 1/200 dilution. Thereafter, cells were washed 3 times with PBS containing 0.3% Triton-X, incubated with secondary antibody and coverslipped with Fluoroshield containing DAPI as a counter stain. Immunofluorescent signals were detected by confocal microscopy.
Immunohistochemistry (Resin sections) analysis of mouse bone tissue cells labeling F-actin with ab205 at 1/200 dilution. Mouse tibia bone samples were fixed in neutral buffered formalin for 72 hours at room temperature, embedded undecalcified in plastic and sectioned at 5 microns. A polyclonal goat anti-mouse Alexa Fluor® 488 secondary antibody was used at 1/700 dilution. The image shows a 1 micron thick z-projection of the chondrocytes in the proximal tibia growth plate.
Blocking: 5% milk for 1 hour at 25°C.
Incubation with primary antibody for 15 hours at 4°C in 5%milk+ PBS+0.1% Tween 20.
Immunocytochemistry/ Immunofluorescence analysis of mouse cortical neurons labeling F-actin with ab205 at 1/100 dilution. The cells were fixed with Ethanol and permeabilized with 0.2% Triton X-100. A polyclonal goat anti-mouse Cy3 conjugated secondary antibody was used at 1/200 dilution.
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