Alexa Fluor® 647 Anti-Iba1 antibody [EPR16588] - Microglia marker

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Alexa Fluor® 647 Anti-Iba1 antibody [EPR16588] - Microglia marker (AB225261)

IHC image of Iba1 staining in a section of formalin-fixed paraffin-embedded normal human tonsil*.

The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6) in a Biocare Medical NxGen pressure cooker using retrieval settings of 110^oC for 20 minutes. Non-specific protein-protein interactions were then blocked in TBS containing 0.025% (v/v) Triton X-100, 0.3M (w/v) glycine and 1% (w/v) BSA for 1h at room temperature. The section was then incubated overnight at +4°C in TBS containing 0.025% (v/v) Triton X-100 and 1% (w/v) BSA with ab225261 at 1/100 dilution (shown in red) and counterstained using ab195887, Mouse monoclonal to alpha Tubulin (Alexa Fluor^® 488), at 1/250 dilution (shown in green). Nuclear DNA was labelled with DAPI (shown in blue). The section was then mounted using Fluoromount^®.

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.

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

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Alexa Fluor® 647 Anti-Iba1 antibody [EPR16588] - Microglia marker (AB225261)

Immunofluorescence staining of Iba1 staining in a section of formalin-fixed paraffin-embedded human Alzheimer’s brain*.

Performed on a Leica BOND. The section was pre-treated using heat mediated antigen retrieval with EDTA (Ph9.0) using retrieval settings of 100°C for 40 minutes. The section was then incubated at room temperature for 1 hour with ab225261 at 1/500 dilution (shown in magenta). Nuclear DNA was labelled with DAPI (shown in blue). The section was then mounted using Dako Fluorescence Mounting Medium ^®.

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.

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

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Alexa Fluor® 647 Anti-Iba1 antibody [EPR16588] - Microglia marker (AB225261)

Image A : composite multiplex immunohistochemistry staining using conjugates panel composed of anti- GFAP (Alexa Fluor^® 488) (ab194324, 1/100 dilution, 5 µg/mL) (green), anti-MOG (Alexa Fluor^® 594) (ab305372, 1/100 dilution, 5 µg/mL) (magenta) and anti-Iba1 (Alexa Fluor^® 647) (ab225261, 1/100 dilution, 5 µg/mL) (white) on Formalin/PFA-fixed paraffin-embedded sections of rat cerebellum. Nuclear DNA was labelled with DAPI (blue).

Image B : Formalin/PFA-fixed paraffin-embedded section of rat cerebellum stained with anti- GFAP (Alexa Fluor^® 488) (ab194324, 1/100 dilution, 5 µg/mL).

Image C : Formalin/PFA-fixed paraffin-embedded section of rat cerebellum stained with anti- MOG (Alexa Fluor^® 594) (ab305372, 1/100 dilution, 5 µg/mL).

Image D : Formalin/PFA-fixed paraffin-embedded section of rat cerebellum stained with anti- Iba1 (Alexa Fluor^® 647) (ab225261, 1/100 dilution, 5 µg/mL).

Immunohistochemistry staining was performed on a Leica Biosystems BOND^® RX instrument. Heat mediated antigen retrieval with EDTA (pH 9.0, 110°C) for 40 mins. The section was then incubated at room temperature for 1 hour and mounted using Dako Fluorescence Mounting Medium^®.

Image acquisition was performed with Leica SP8 confocal microscope. For other IHC staining systems (automated and non-automated), customers should optimize variable parameters such as antigen retrieval conditions, antibody concentrations and incubation times.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Alexa Fluor® 647 Anti-Iba1 antibody [EPR16588] - Microglia marker (AB225261)

Immunofluorescence staining of Iba1 staining in a section of formalin-fixed paraffin-embedded normal human cerebellum*.

Performed on a Leica BOND. The section was pre-treated using heat mediated antigen retrieval with EDTA (Ph9.0) using retrieval settings of 100°C for 40 minutes. The section was then incubated at room temperature for 1 hour with ab225261 at 1/500 dilution (shown in magenta). Nuclear DNA was labelled with DAPI (shown in blue). The section was then mounted using Dako Fluorescence Mounting Medium ^®.

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.

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