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Our Abpromise guarantee covers the use of ab89780 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|Flow Cyt||Use 1µg for 106 cells. ab170190-Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.|
|IHC-P||1/2000 - 1/4000. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.|
|WB||Use a concentration of 1 - 5 µg/ml. Predicted molecular weight: 63 kDa. Isoform a, 69 kDa; Isoform b, 63 kDa.|
|ELISA||Use at an assay dependent concentration.|
ab89780 staining MAG in lamb spinal cord tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with formaldehyde and blocked with 1% BSA for 10 minutes at 21°C; antigen retrieval was by heat mediation in a citrate buffer. Samples were incubated with primary antibody (1/5000 in blocking buffer) for 2 hours at 21°C. A Biotin-conjugated Goat anti-mouse IgG polyclonal (1/200) was used as the secondary antibody.
ab89780 stained SKNSH cells. The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody ab89780 at 10µg/ml overnight at +4°C. The secondary antibody (green) was DyLight® 488 goat anti- mouse (ab96879) IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.
ab89780 staining MAG in Mouse brain tissue sections by Immunohistochemistry (PFA perfusion fixed frozen sections). Tissue samples were fixed by perfusion with acetone, permeablized with methanol and blocked with 5% BSA for 1 hour at 37°C. The sample was incubated with primary antibody (1/100) at 4°C for 18 hours. An Alexa Fluor® 594-conjugated Goat anti-mouse polyclonal (1/200) was used as the secondary antibody.
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