Anti-Iba1 antibody [EPR16588] - Microglia marker – Chicken IgY (Chimeric)

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-Iba1 antibody [EPR16588] - Microglia marker – Chicken IgY (Chimeric) (AB318302)

This image was produced using ab317805, the same antibody clone but in a different formulation. ab318305 staining Iba1 in THP-1 (positive) and HeLa (negative) cells. The cells were fixed with 100% methanol (5 min), permeabilized with 0.1% PBS-Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1%PBS-Tween for 1h. The cells were then incubated overnight at 4°C with ab318305 at 5 µg/ml and ab202272, Alexa Fluor® 594 Rabbit monoclonal [EP1332Y] to alpha Tubulin - Microtubule Marker (shown in pseudocolour magenta). Cells were then incubated with ab150173, Goat Anti-Chicken IgY H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution (shown in green). Nuclear DNA was labelled with DAPI (shown in blue). Image was acquired with a confocal microscope (Leica-Microsystems TCS SP8) and a single confocal section is shown.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Iba1 antibody [EPR16588] - Microglia marker – Chicken IgY (Chimeric) (AB318302)

This image was produced using ab317805, the same antibody clone but in a different formulation. Immunofluorescence staining of Iba1 in a section of formalin-fixed paraffin-embedded human brain. Performed on a Leica BOND. The section was pre-treated using heat mediated antigen retrieval with sodium citrate (pH6.0) for 20 minutes. The section was then incubated at room temperature for 1 hour with ab318305 at 1 μg/ml, and then incubated for 1 hour with ab150173, Goat Anti-Chicken IgY H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution (shown in green). 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.

• IHC-Fr

Lab

Immunohistochemistry (Frozen sections) - Anti-Iba1 antibody [EPR16588] - Microglia marker – Chicken IgY (Chimeric) (AB318302)

Immunohistochemical analysis of 4% PFA, 0.2% Triton X-100 permeabilized frozen Mouse cerebrum (perfused-fixed) tissue labelling Iba1 with ab318302 at 1/50 dilution followed by Goat Anti-Chicken IgY H&L (Alexa Fluor® 488) preadsorbed (ab150173) at 1/1000 dilution (Green).

Confocal image showing positive staining on mouse cerebrum. The nuclear counterstain was DAPI (Blue). The section was incubated with ab318302 for 60 mins at room temperature. The section was then mounted using Fluoromount®.The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

Secondary antibody control : Secondary antibody is Goat Anti-Chicken IgY H&L (Alexa Fluor® 488) preadsorbed ab150173 at 1/1000.

Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20)

• IHC-Fr

Lab

Immunohistochemistry (Frozen sections) - Anti-Iba1 antibody [EPR16588] - Microglia marker – Chicken IgY (Chimeric) (AB318302)

Immunohistochemical analysis of 4% PFA, 0.2% Triton X-100 permeabilized frozen Rat cerebrum (perfused-fixed) tissue labelling Iba1 with ab318302 at 1/50 dilution followed by Goat Anti-Chicken IgY H&L (Alexa Fluor® 488) preadsorbed (ab150173) at 1/1000 dilution (Green).

Confocal image showing positive staining on Rat cerebrum. The nuclear counterstain was DAPI (Blue). The section was incubated with ab318302 for 60 mins at room temperature. The section was then mounted using Fluoromount®.The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

Secondary antibody control : Secondary antibody is Goat Anti-Chicken IgY H&L (Alexa Fluor® 488) preadsorbed ab150173 at 1/1000.

Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20)

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Iba1 antibody [EPR16588] - Microglia marker – Chicken IgY (Chimeric) (AB318302)

This image was produced using ab317805, the same antibody clone but in a different formulation. Immunofluorescence staining of Iba1 in a section of formalin-fixed paraffin-embedded mouse brain. Performed on a Leica BOND. The section was pre-treated using heat mediated antigen retrieval with sodium citrate (pH6.0) for 20 minutes. The section was then incubated at room temperature for 1 hour with ab318305 at 1 μg/ml, and then incubated for 1 hour with ab150173, Goat Anti-Chicken IgY H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution (shown in green). 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.

• IHC-Fr

Lab

Immunohistochemistry (Frozen sections) - Anti-Iba1 antibody [EPR16588] - Microglia marker – Chicken IgY (Chimeric) (AB318302)

Immunohistochemical analysis of 4% PFA, 0.2% Triton X-100 permeabilized frozen Rat liver (perfused-fixed) tissue labelling Iba1 with ab318302 at 1/50 dilution followed by Goat Anti-Chicken IgY H&L (Alexa Fluor® 488) preadsorbed (ab150173) at 1/1000 dilution (Green).

Confocal image showing positive staining on Rat liver. The nuclear counterstain was DAPI (Blue). The section was incubated with ab318302 for 60 mins at room temperature. The section was then mounted using Fluoromount®.The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

Secondary antibody control : Secondary antibody is Goat Anti-Chicken IgY H&L (Alexa Fluor® 488) preadsorbed ab150173 at 1/1000.

Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20)

• IHC-Fr

Lab

Immunohistochemistry (Frozen sections) - Anti-Iba1 antibody [EPR16588] - Microglia marker – Chicken IgY (Chimeric) (AB318302)

Immunohistochemical analysis of 4% PFA, 0.2% Tritin X-100 permeabilized frozen Mouse liver tissue labelling Iba1 with ab318302 at 1/50 dilution Goat Anti-Chicken IgY H&L (Alexa Fluor® 488) preadsorbed (ab150173) at 1/1000 dilution (Green)

Secondary antibody control : Secondary antibody is Goat Anti-Chicken IgY H&L (Alexa Fluor® 488) preadsorbed (ab150173) at 1/1000 dilution

Confocal image showing positive staining on mouse liver. The nuclear counterstain was DAPI (Blue). The section was incubated with ab318302 for 60 mins at room temperature. The section was then mounted using Fluoromount®.The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20)