Anti-Iba1 antibody [EPR16588] - Microglia marker - Rat IgG2a (Chimeric)

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Iba1 antibody [EPR16588] - Microglia marker - Rat IgG2a (Chimeric) (AB300156)

Immunohistochemical analysis of paraffin-embedded Human tonsil tissue labeling Iba1 with ab300156 at 1/10000 (0.108 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on human tonsil. The section was incubated with ab300156 for 30 mins at room temperature, followed by anti-Rat IgG antibody (ab102248) for 8 mins during the LeicaDS9800 kit staining procedure. The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin. Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-Iba1 antibody [EPR16588] - Microglia marker - Rat IgG2a (Chimeric) (AB300156)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized MCF7 (Human breast adenocarcinoma epithelial cell) / THP-1 (Human monocytic leukemia monocyte) cells labelling Iba1 with ab300156 at 1/1000 dilution (0.1ug) (Red) compared with a Rat monoclonal IgG (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat Anti-Rat IgG Fc (DyLight 488, ab96971) at 1/2000 dilution was used as the secondary antibody. Negative control : MCF7 (PMID : 30386176)

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Iba1 antibody [EPR16588] - Microglia marker - Rat IgG2a (Chimeric) (AB300156)

Immunohistochemical analysis of paraffin-embedded Human liver tissue labeling Iba1 with ab300156 at 1/10000 (0.108 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on Kupffer cells in the human liver. The section was incubated with ab300156 for 30 mins at room temperature, followed by anti-Rat IgG antibody (ab102248) for 8 mins during the LeicaDS9800 kit staining procedure. The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin. Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Iba1 antibody [EPR16588] - Microglia marker - Rat IgG2a (Chimeric) (AB300156)

Composite multiplex immunofluorescence staining of Iba1, GFAP and MAP2 staining in a section of formalin-fixed paraffin-embedded human cerebral cortex*.

Performed on a Leica BOND. The section was pre-treated using heat mediated antigen retrieval with Sodium citrate (Ph6.0) using retrieval settings of 100°C for 20 minutes. The section was then incubated at room temperature for 1 hour with ab300156 at 1µg/ml dilution (shown in green), ab183830 at 1µg/ml (shown in magenta), and ab302644 at 1µg/ml (shown in yellow). Then incubated for 1 hour with ab150161 Goat F(ab')2 Anti-Rat IgG Fc (Alexa Fluor® 488) preadsorbed 1/1000, ab150083 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 647) preadsorbed 1/1000, and ab150134 Donkey Anti-Goat IgG H&L (Alexa Fluor® 555) preadsorbed 1/1000. Nuclear DNA was labelled with DAPI (shown in blue). The section was then mounted using Dako Fluorescence Mounting Medium ^®.

Image was taken with the EVOS™ S1000 Spatial Imaging System (ThermoFisher Scientific) with spectral unmixing and minor subsequent contrast adjustment.

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

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Iba1 antibody [EPR16588] - Microglia marker - Rat IgG2a (Chimeric) (AB300156)

Immunohistochemical analysis of paraffin-embedded Human cerebrum tissue labeling Iba1 with ab300156 at 1/10000 (0.108 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on microglia in the human cerebrum. The section was incubated with ab300156 for 30 mins at room temperature, followed by anti-Rat IgG antibody (ab102248) for 8 mins during the LeicaDS9800 kit staining procedure. The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin. Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Iba1 antibody [EPR16588] - Microglia marker - Rat IgG2a (Chimeric) (AB300156)

Immunohistochemical analysis of paraffin-embedded Rat cerebrum tissue labeling Iba1 with ab300156 at 1/10000 (0.108 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on microglia in the rat cerebrum. The section was incubated with ab300156 for 30 mins at room temperature, followed by anti-Rat IgG antibody (ab102248) for 8 mins during the LeicaDS9800 kit staining procedure. The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin. Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Iba1 antibody [EPR16588] - Microglia marker - Rat IgG2a (Chimeric) (AB300156)

Immunohistochemical analysis of paraffin-embedded Mouse cerebrum tissue labeling Iba1 with ab300156 at 1/10000 (0.108 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on microglia in the mouse cerebrum. The section was incubated with ab300156 for 30 mins at room temperature, followed by anti-Rat IgG antibody (ab102248) for 8 mins during the LeicaDS9800 kit staining procedure. The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin. Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Iba1 antibody [EPR16588] - Microglia marker - Rat IgG2a (Chimeric) (AB300156)

Immunohistochemical analysis of paraffin-embedded Mouse liver tissue labeling Iba1 with ab300156 at 1/10000 (0.108 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on Kupffer cells in the mouse liver. The section was incubated with ab300156 for 30 mins at room temperature, followed by anti-Rat IgG antibody (ab102248) for 8 mins during the LeicaDS9800 kit staining procedure. The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin. Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Iba1 antibody [EPR16588] - Microglia marker - Rat IgG2a (Chimeric) (AB300156)

Immunohistochemical analysis of paraffin-embedded Rat liver tissue labeling Iba1 with ab300156 at 1/10000 (0.108 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on Kupffer cells in the rat liver. The section was incubated with ab300156 for 30 mins at room temperature, followed by anti-Rat IgG antibody (ab102248) for 8 mins during the LeicaDS9800 kit staining procedure. The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin. Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins