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

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

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.

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.

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.