Anti-CD11b antibody [EPR1344] (ab133357)

Formaldehyde-fixed, paraffin-embedded rat bone marrow tissue stained for CD11b using ab133357 at 1/5000 in immunohistochemical analysis.

Heat mediated antigen retrieval with EDTA buffer pH 9 was performed before commencing with staining protocol. 1% casein was used as blocking agent.

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IHC image of CD11b staining in a formalin fixed, paraffin embedded human normal spleen tissue section*, performed on a Leica Bond™ system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab133357 at 1/4000 dilution, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

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.

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

Blocking and diluting buffer: 5% NFDM/TBST

Immunohistochemical analysis of CD11b in paraffin embedded human spleen tissue, using unpurified ab133357 at a dilution of 1/100.

Immunohistochemical analysis of CD11b in paraffin embedded human tonsil tissue, using unpurified ab133357 at a dilution of 1/100.

Blocking buffer: 5% NFDM/TBST

Dilution buffer: 5% NFDM/TBST

Blocking buffer: 5% NFDM/TBST

Dilution buffer: 5% NFDM/TBST

Immunohistochemical staining of paraffin embedded human spleen with purified ab133357 at a working dilution of 1 in 4000. The secondary antibody used is a HRP goat anti-rabbit (ab97051). The sample is counter-stained with hematoxylin. Antigen retrieval was perfomed using Tris-EDTA buffer, pH 9.0. PBS was used instead of the primary antibody as the negative control, and is shown in the inset.