Anti-Myelin Basic Protein antibody - Oligodendrocyte Marker

• ICC

Lab

Immunocytochemistry - Anti-Myelin Basic Protein antibody - Oligodendrocyte Marker (AB40390)

ab40390 staining Myelin Basic Protein in primary hippocampal rat neurons/glia, (obtained from Neuromics, cat. no. PC35101), DIV14. The cells were fixed with 100% methanol (5 min), permeabilized with 0.1% PBS-Tween for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1%PBS-Tween for 1h. The cells were then incubated overnight at 4°C with ab40390 at 1μg/ml and ab7291, Mouse monoclonal [DM1A] to alpha Tubulin - Loading Control. Cells were then incubated with ab150081, Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor^® 488), pre-adsorbed at 1/1000 dilution (shown in green) and ab150120, Goat polyclonal Secondary Antibody to Mouse IgG - H&L (Alexa Fluor^® 594), pre-adsorbed at 1/1000 dilution (shown in pseudocolour red). Nuclear DNA was labelled with DAPI (shown in blue).

Also suitable in cells fixed with 4% paraformaldehyde (10 min).

Image was acquired with a confocal microscope (Leica-Microsystems TCS SP8) and a single confocal section is shown.

• ICC

Unknown

Immunocytochemistry - Anti-Myelin Basic Protein antibody - Oligodendrocyte Marker (AB40390)

ICC/IF image of ab40390 stained PC12 cells. The cells were 100% methanol fixed (5 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 (ab40390, 5µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit 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).

• WB

Project1449****

Western blot - Anti-Myelin Basic Protein antibody - Oligodendrocyte Marker (AB40390)

This antibody was raised against an immunogen that is predicted to recognize isoforms (5, 7, 8, 10 and 13) of Myelin Basic Protein (MBP). The predicted molecular weights of isoforms (5, 7, 8, 10 and 13) are 18.5kDa, 17kDa, 14kDa, 21kDa and 13kDa respectively.

All lanes:

Western blot - Anti-Myelin Basic Protein antibody - Oligodendrocyte Marker (ab40390) at 1 µg/mL

Lane 1:

Brain (Mouse) Tissue Lysate at 10 µg

Lane 2:

Brain (Rat) Tissue Lysate - normal tissue at 10 µg

Secondary

All lanes:

Goat polyclonal to rabbit IgG - H&L - Pre adsorbed (HRP) at 1/3000 dilution

Predicted band size: 33 kDa

Observed band size: 18 kDa,23 kDa,45 kDa

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