Anti-Myelin Basic Protein antibody [IGX3421] - Oligodendrocyte Marker

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-Myelin Basic Protein antibody [IGX3421] - Oligodendrocyte Marker (AB209328)

ab209328 staining Myelin Basic Protein in SK-N-SH (Human neuroblastoma cell line) cells. The cells were fixed with 4% formaldehyde (10 minutes), permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal donkey serum/0.3M glycine in 0.1% PBS-Tween for 1 hour. The cells were then incubated overnight at +4°C with ab209328 at a 5 μg/ml concentration, then detected with a donkey anti-human (Alexa Fluor^® 488) secondary antibody at a 1/2000 dilution (shown in green). Nuclear DNA was labelled with DAPI (shown in blue), and ab195889, Mouse monoclonal to alpha Tubulin (Alexa Fluor^® 594), at a 1/250 dilution (shown in red).

Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

• ICC

Lab

Immunocytochemistry - Anti-Myelin Basic Protein antibody [IGX3421] - Oligodendrocyte Marker (AB209328)

Immunocytochemistry analysis of 4% Paraformaldehyde-fixed 0.1% TritonX-100 permeabilized SK-N-SH (human neuroblastoma epithelial cell) cells staining with ab209328 at 1/100 dilution. Counterstained with ab206369 Anti-beta Tubulin rabbit monoclonal antibody (Alexa Fluor® 594) at 1/1000 dilution. The secondary used was Goat anti-Human IgG (H+L) Cross-Adsorbed Secondary Antibody (Alexa Fluor® 488) at 1/1000 dilution. Confocal image showing cytoplasmic and weak nuclear staining in SK-N-SH cell line (shown in green). The counterstain was observed in red. Nuclear DNA was labelled with DAPI (shown in blue). Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8). Image was taken with a confocal microscope(Leica-Microsystems, TCS SP8).

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Myelin Basic Protein antibody [IGX3421] - Oligodendrocyte Marker (AB209328)

IHC image of Myelin Basic Protein staining in a section of formalin-fixed paraffin-embedded normal human hippocampus and normal human pancreas*, performed on a Leica BOND^TM. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6, epitope retrieval solution 1) for 20 minutes. The section was then incubated with ab209328, 1/1000 dilution, for 15 minutes at room temperature.

An HRP-conjugated goat anti-Human IgG secondary was used for 15 minutes at room temperature. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX. The inset secondary-only control image is taken from an identical assay without primary antibody.

For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

*Human pancreas tissue was obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-Myelin Basic Protein antibody [IGX3421] - Oligodendrocyte Marker (AB209328)

ab209328 staining Myelin Basic Protein in SHSY5Y cells. The cells were fixed with 100% methanol (5 minutes), permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal donkey serum/0.3M glycine in 0.1% PBS-Tween for 1 hour. The cells were then incubated overnight at +4°C with ab209328 at a 5 μg/ml concentration, then detected with a donkey anti-human (Alexa Fluor^® 488) secondary antibody at a 1/1000 dilution (shown in green). Nuclear DNA was labelled with DAPI (shown in blue), and ab195884, Rat monoclonal to alpha Tubulin (Alexa Fluor^® 647), at a 1/250 dilution (shown in red).

Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Myelin Basic Protein antibody [IGX3421] - Oligodendrocyte Marker (AB209328)

IHC image of Myelin Basic Protein staining in a section of formalin-fixed paraffin-embedded normal mouse brain and normal mouse pancreas, performed on a Leica BOND^TM. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6, epitope retrieval solution 1) for 20 minutes. The section was then incubated with ab209328, 1/1000 dilution, for 15 minutes at room temperature.

An HRP-conjugated goat anti-Human IgG secondary was used for 15 minutes at room temperature. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX. The inset secondary-only control image is taken from an identical assay without primary antibody.

For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Myelin Basic Protein antibody [IGX3421] - Oligodendrocyte Marker (AB209328)

IHC image of Myelin Basic Protein staining in a section of formalin-fixed paraffin-embedded normal rat brain and normal rat pancreas, performed on a Leica BOND^TM. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6, epitope retrieval solution 1) for 20 minutes. The section was then incubated with ab209328, 1/1000 dilution, for 15 minutes at room temperature.

An HRP-conjugated goat anti-Human IgG secondary was used for 15 minutes at room temperature. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX. The inset secondary-only control image is taken from an identical assay without primary antibody.

For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

• ICC

Lab

Immunocytochemistry - Anti-Myelin Basic Protein antibody [IGX3421] - Oligodendrocyte Marker (AB209328)

Immunocytochemistry analysis of 4% Paraformaldehyde-fixed 0.1% TritonX-100 permeabilized C6 (rat glial tumor glial cell) cells staining with ab209328 at 1/100 dilution. Counterstained with ab206369 Anti-beta Tubulin rabbit monoclonal antibody (Alexa Fluor® 594) at 1/400 dilution. The secondary used was Goat anti-Human IgG (H+L) Cross-Adsorbed Secondary Antibody (Alexa Fluor® 488) at 1/1000 dilution. Confocal image showing cytoplasmic and weak nuclear staining in C6 cell line (shown in green). The counterstain was observed in red. Nuclear DNA was labelled with DAPI (shown in blue). Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

• WB

Lab

Western blot - Anti-Myelin Basic Protein antibody [IGX3421] - Oligodendrocyte Marker (AB209328)

Exposure time :

Lane 1 : 30 seconds.

Lanes 2-3 : 2 minutes.

Lane 4 : 8 minutes.

This blot was produced using a 4-12% Bis-tris gel under the MES buffer system. The gel was run at 200V for 35 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 3% milk before being incubated with ab209328 overnight at 4°C. Antibody binding was visualised using ECL development solution ab133406.

All lanes:

Western blot - Anti-Myelin Basic Protein antibody [IGX3421] - Oligodendrocyte Marker (ab209328) at 0.25 µg/mL

Lane 1:

Human brain tissue lysate - total protein (<a href='/en-us/products/unavailable/human-brain-tissue-lysate-total-protein-ab29466'>ab29466</a>) at 10 µg

Lane 2:

Mouse brain tissue lysate at 10 µg

Lane 3:

Rat brain tissue lysate at 10 µg

Lane 4:

Myelin Basic Protein (Recombinant protein) at 0.1 µg

Secondary

All lanes:

HRP conjugated Goat Anti-Human IgG (H+L) at 1/10000 dilution

Predicted band size: 33 kDa

Observed band size: 18 kDa,23 kDa,24 kDa

true

• WB

Lab

Western blot - Anti-Myelin Basic Protein antibody [IGX3421] - Oligodendrocyte Marker (AB209328)

This blot was produced using a 4-12% Bis-tris gel under the MES buffer system. The gel was run at 200V for 35 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 3% milk before being incubated with ab209328 overnight at 4°C. Antibody binding was visualised using ECL development solution ab133406.

All lanes:

Western blot - Anti-Myelin Basic Protein antibody [IGX3421] - Oligodendrocyte Marker (ab209328) at 1 µg/mL

All lanes:

Mouse brain tissue lysate at 10 µg

Secondary

All lanes:

HRP conjugated Goat Anti-Human IgG (H+L) at 1/10000 dilution

Predicted band size: 33 kDa

Observed band size: 17 kDa,20 kDa

true

Exposure time: 2min

• ELISA

Lab

ELISA - Anti-Myelin Basic Protein antibody [IGX3421] - Oligodendrocyte Marker (AB209328)

ELISA using ab209328 for 16 hours at 4°C. ab7153 goat anti human was used as a secondary at a 1/5000 dilution for 1 hour at Room Temperature.