Microglia Marker (CD11b, CD45, Iba1, TMEM119) Antibody Sampler Panel (ab226482)

IHC image of TMEM119 staining in a section of frozen normal mouse brain. No antigen retrieval step was performed prior to staining. Non-specific protein-protein interactions were then blocked in PBS containing 0.5% (v/v) Triton X-100, 0.3 M (w/v) glycine and 1% (w/v) BSA for 1 h at room temperature. The section was then incubated overnight at +4°C in PBS containing 0.5% (v/v) Triton X-100 and 1% (w/v) BSA with ab209064 at 0.5 µg/ml. The secondary antibody was ab150087 (shown in red) used at 2 µg/ml for 1 hour at room temperature. Nuclear DNA was labelled with DAPI (shown in blue). The section was then mounted using Fluoromount^®. The image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

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

Immunohistochemical staining of paraffin embedded human tonsil with purified ab40763 at a working dilution of 1/250. The secondary antibody used is HRP goat anti-rabbit IgG H&L (ab97051) at 1/500. 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.

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.

IHC image of Iba1 staining in mouse normal brain formalin fixed paraffin embedded tissue section, performed on a Leica Bond™ system using the standard protocol B. 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 ab178846, 1/2000 dilution, for 15 mins at room temperature. A goat anti-rabbit biotinylated secondary antibody was used to detect the primary, and visualized using an HRP conjugated ABC 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.