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 sections were pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20mins. The section was then incubated with ab216668, 1ug/ml, for 15 mins at room temperature.

An HRP-conjugated goat anti-Rabbit IgG secondary (ab97080) was used for 15 mins 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.

Pancreatic tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre.

IHC image of Myelin Basic Protein staining in a section of formalin-fixed paraffin-embedded normal human brain and normal human pancreas, performed on a Leica BOND^TM. The sections were pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20mins. The section was then incubated with ab216668, 1ug/ml, for 15 mins at room temperature.

An HRP-conjugated goat anti-Rabbit IgG secondary (ab97080) was used for 15 mins 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.

Pancreatic tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre.

Ab216668 staining myelin basic protein in Mouse brain tissue sections by Immunohistochemistry (IHC-P - formaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with formaldehyde and blocked with 2% BSA for 10 minutes at 21°C; antigen retrieval was by heat mediation in a citrate buffer. Samples were incubated with primary antibody (1/4000 in blocking buffer) for 2 hours at 21°C. A diluted Biotin-conjugated Goat anti-rabbit IgG polyclonal (1/300) was used as the secondary antibody.

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All lanes : Anti-Myelin Basic Protein antibody [IGX3421R-1] (ab216668) at 0.25 µg/ml

Lane 1 : Human brain tissue lysate - total protein (ab29466) 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 : Goat Anti-Rabbit IgG H&L (HRP) preadsorbed at 1/50000 dilution

Developed using the ECL technique.

Performed under reducing conditions.

Predicted band size: 33 kDa
Observed band size: 17,20,23 kDa why is the actual band size different from the predicted?

Exposure time :

Lane 1 & 4 : 90 seconds.

Lanes 2 & 3 : 4 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 ab216668 overnight at 4°C. Antibody binding was visualised using ECL development solution ab133406.

Ab216668 staining myelin basic protein in Rat brain tissue sections by Immunohistochemistry (IHC-P - formaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with formaldehyde and blocked with 2% BSA for 10 minutes at 21°C; antigen retrieval was by heat mediation in a citrate buffer. Samples were incubated with primary antibody (1/6000 in blocking buffer) for 2 hours at 21°C. A diluted Biotin conjugated Goat anti- rabbit IgG polyclonal (1/300) was used as the secondary antibody.

See Abreview