Western blot - Anti-Myelin Basic Protein antibody [12] (ab7349)
The molecular weight observed was in consistency with the literature (PMID: 9299539).
Blocking buffer: 5% NFDM/TBST.
All lanes:
Western blot - Anti-Myelin Basic Protein antibody [12] (ab7349) at 1/5000 dilution
All lanes:
Human cerebellum tissue lysate at 10 µg
Secondary
All lanes:
Western blot - Goat Anti-Rat IgG H&L (HRP) (ab205720) at 1/10000 dilution
Developed using the ECL technique.
Predicted band size: 33 kDa
Observed band size: 18.5 kDa
Exposure time: 1s
Western blot - Anti-Myelin Basic Protein antibody [12] (ab7349)
The molecular weight observed was in consistency with the literature (PMID: 9299539).
Blocking buffer: 5% NFDM/TBST.
Exposure time: Lane 1: 3 seconds; Lane 2: 1 second.
All lanes:
Western blot - Anti-Myelin Basic Protein antibody [12] (ab7349) at 1/5000 dilution
Lane 1:
Mouse brain tissue lysate at 10 µg
Lane 2:
Rat brain tissue lysate at 10 µg
Secondary
All lanes:
Western blot - Goat Anti-Rat IgG H&L (HRP) (ab205720) at 1/10000 dilution
Predicted band size: 33 kDa
Observed band size: 14-21.5 kDa
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Myelin Basic Protein antibody [12] (ab7349)
Immunohistochemical analysis of paraffin-embedded Human cerebrum labeling Myelin Basic Protein with ab7349 at 1/4000 dilution (0.259 μg/ml) followed by a ready to use Goat Anti-Rat IgG H&L (HRP polymer; ab214882). The section was incubated with ab7349 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.
Heat mediated antigen retrieval with Citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 mins.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Myelin Basic Protein antibody [12] (ab7349)
Immunohistochemical analysis of paraffin-embedded Human cerebrum labeling Myelin Basic Protein with ab7349 at 1/4000 dilution (0.259 μg/ml) followed by a ready to use Goat Anti-Rat IgG H&L (HRP polymer; ab214882). The section was incubated with ab7349 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with DAPI.
Heat mediated antigen retrieval with Citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 mins.
Immunocytochemistry/ Immunofluorescence - Anti-Myelin Basic Protein antibody [12] (ab7349)
Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized mouse primary neural/glia cells labelling Myelin Basic Protein with ab7349 at a 1/200 dilution, followed by Goat anti-Rat (Alexa Fluor® 488) (ab150157) secondary antibody at a 1/1000 dilution. Confocal image showing positive staining in mouse primary oligodendroglia cells (shown in green). Nuclear DNA was labeled with DAPI (shown in blue).
Counterstained with Anti-MAP2 antibody (ab183830) at a 1/1000 dilution, followed by Goat Anti-Rabbit secondary (Alexa Fluor® 594) (ab150080) at a 1/1000 dilution (shown in magenta).
Confocal scanning Z step was set as 0.3 μm followed by image processing with maximum Z projection. Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).
The negative controls are as follows:
-ve control 1: Myelin Basic Protein (ab7349) at a 1/200 dilution, followed by Goat anti-Rabbit (Alexa Fluor® 594) (ab150080) at a 1/1000 dilution.
-ve control 2: Anti-MAP2 (ab183830) at a 1/1000 dilution, followed by Goat anti-Rat (Alexa Fluor® 488) (ab150157) at a 1/1000 dilution.
