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

• IHC-P

Lab

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

Immunohistochemical analysis of paraffin-embedded Human cerebrum tissue labelling Iba1 with ab318302 at 1/5000 dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on human cerebrum.
The primary antibody was incubated for 30 mins at room temperature, followed by anti-Chicken IgY antibody (ab97136) 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 was performed 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 – Chicken IgY (Chimeric) (AB318302)

Immunohistochemical analysis of paraffin-embedded Human liver tissue labelling Iba1 with ab318302 at 1/5000 dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on human liver.
The primary antibody was incubated for 30 mins at room temperature, followed by anti-Chicken IgY antibody (ab97136) 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 was performed 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 – Chicken IgY (Chimeric) (AB318302)

Immunohistochemical analysis of paraffin-embedded Human tonsil tissue labelling Iba1 with ab318302 at 1/5000 dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on human tonsil.
The primary antibody was incubated for 30 mins at room temperature, followed by anti-Chicken IgY antibody (ab97136) 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 was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Anti-Iba1 antibody [EPR16588] - Microglia marker – Chicken IgY (Chimeric) (AB318302)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized MCF7 (human breast adenocarcinoma epithelial cell, Left) / THP-1 (human monocytic leukemia monocyte, Right) cells labelling Iba1 with ab318302 at 1/5000 dilution (0.01ug)/ Red compared with a purified Chicken IgY Isotype Control Antibody /Black and cell without incubation with primary antibody and secondary antibody / Blue.

Goat Anti-Chicken IgY H&L (Alexa Fluor^® 488) preadsorbed (ab150173) at 1/5000 dilution was used as the secondary antibody.

Negative control : MCF7

• 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-P

Lab

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

Immunohistochemical analysis of paraffin-embedded Rat cerebrum tissue labelling Iba1 with ab318302 at 1/5000 dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on rat cerebrum.
The primary antibody was incubated for 30 mins at room temperature, followed by anti-Chicken IgY antibody (ab97136) 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 was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• 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-P

Lab

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

Immunohistochemical analysis of paraffin-embedded Mouse cerebrum tissue labelling Iba1 with ab318302 at 1/5000 dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on mouse cerebrum.
The primary antibody was incubated for 30 mins at room temperature, followed by anti-Chicken IgY antibody (ab97136) 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 was performed 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 – Chicken IgY (Chimeric) (AB318302)

Immunohistochemical analysis of paraffin-embedded Mouse liver tissue labelling Iba1 with ab318302 at 1/5000 dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on mouse liver.
The primary antibody was incubated for 30 mins at room temperature, followed by anti-Chicken IgY antibody (ab97136) 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 was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• ICC/IF

Lab

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

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized mouse primary neural/glia cell cells labelling Iba1 with ab318302 at 1/200 dilution (Green).

Confocal image showing microglial staining in mouse primary neural/glia cell (shown in green). The counterstain was observed in magenta. Nuclear DNA was labelled with DAPI (shown in blue). Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).
Confocal scanning Z step was set as 0.3 µm followed by image processing with maximum Z projection.

Rabbit anti-Iba1 antibody was used to counterstain Iba1 at 1/1000 dilution (Magenta). The Nuclear counterstain was DAPI (Blue).

• ICC/IF

Lab

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

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized rat primary neural/glia cell cells labelling Iba1 with ab318302 at 1/200 dilution (Green).

Confocal image showing microglial staining in rat primary neural/glia cell (shown in green). The counterstain was observed in magenta. Nuclear DNA was labelled with DAPI (shown in blue). Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).
Confocal scanning Z step was set as 0.3 µm followed by image processing with maximum Z projection.

Rabbit anti-Iba1 antibody was used to counterstain Iba1 at 1/1000 dilution (Magenta). The Nuclear counterstain was DAPI (Blue).

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Anti-Iba1 antibody [EPR16588] - Microglia marker – Chicken IgY (Chimeric) (AB318302)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized Mouse primary neural/glia cells labelling Iba1 with ab318302 at 1/50 dilution (1ug)/ Right compared with a purified Chicken IgY Isotype Control Antibody /Left.

Goat Anti-Chicken IgY H&L (Alexa Fluor^® 488) preadsorbed (ab150173) at 1/5000 dilution was used as the secondary antibody.

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Anti-Iba1 antibody [EPR16588] - Microglia marker – Chicken IgY (Chimeric) (AB318302)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized Rat primary neural/glia cells labelling Iba1 with ab318302 at 1/50 dilution (1ug)/ Right compared with a purified Chicken IgY Isotype Control Antibody /Left.

Goat Anti-Chicken IgY H&L (Alexa Fluor^® 488) preadsorbed (ab150173) at 1/5000 dilution was used as the secondary antibody.

• 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 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-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)

• 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)

• WB

Lab

Western blot - Anti-Iba1 antibody [EPR16588] - Microglia marker – Chicken IgY (Chimeric) (AB318302)

Blocking and diluting buffer and concentration : 5% NFDM/TBST

This blot was developed using a high-sensitivity ECL substrate, allowing for the detection of proteins in the mid-femtogram range.

ab181602 was used as a GAPDH loading control.

All lanes:

Western blot - Anti-Iba1 antibody [EPR16588] - Microglia marker – Chicken IgY (Chimeric) (ab318302) at 1/1000 dilution

Lane 1:

Human cerebellum tissue lysate at 20 µg

Lane 2:

Human liver tissue lysate at 20 µg

Lane 3:

Human testis tissue lysate at 20 µg

Lane 4:

Human spleen tissue lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Chicken IgY H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-chicken-igy-h-l-hrp-ab6877'>ab6877</a>) at 1/20000 dilution

Observed band size: 16 kDa

true

Exposure time: 3min

• WB

Lab

Western blot - Anti-Iba1 antibody [EPR16588] - Microglia marker – Chicken IgY (Chimeric) (AB318302)

Blocking and diluting buffer and concentration : 5% NFDM/TBST

This blot was developed using a high-sensitivity ECL substrate, allowing for the detection of proteins in the mid-femtogram range.

ab181602 was used as a GAPDH loading control.

All lanes:

Western blot - Anti-Iba1 antibody [EPR16588] - Microglia marker – Chicken IgY (Chimeric) (ab318302) at 1/1000 dilution

Lane 1:

THP-1 (human monocytic leukemia monocyte) whole cell lysate at 20 µg

Lane 2:

U-937 (human histiocytic lymphoma monocyte) whole cell lysate at 20 µg

Lane 3:

MCF7 (human breast adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lane 4:

293T (human embryonic kidney epithelial cell) whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Chicken IgY H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-chicken-igy-h-l-hrp-ab6877'>ab6877</a>) at 1/20000 dilution

Observed band size: 16 kDa

true

Exposure time: 3min

• WB

Lab

Western blot - Anti-Iba1 antibody [EPR16588] - Microglia marker – Chicken IgY (Chimeric) (AB318302)

Blocking and diluting buffer and concentration : 5% NFDM/TBST

This blot was developed using a high-sensitivity ECL substrate, allowing for the detection of proteins in the mid-femtogram range.

ab181602 was used as a GAPDH loading control.

All lanes:

Western blot - Anti-Iba1 antibody [EPR16588] - Microglia marker – Chicken IgY (Chimeric) (ab318302) at 1/1000 dilution

Lane 1:

Rat brain tissue lysate at 20 µg

Lane 2:

Rat heart tissue lysate at 20 µg

Lane 3:

Rat kidney tissue lysate at 20 µg

Lane 4:

Rat spleen tissue lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Chicken IgY H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-chicken-igy-h-l-hrp-ab6877'>ab6877</a>) at 1/20000 dilution

Observed band size: 16 kDa

true

Exposure time: 3min

• WB

Lab

Western blot - Anti-Iba1 antibody [EPR16588] - Microglia marker – Chicken IgY (Chimeric) (AB318302)

Blocking and diluting buffer and concentration : 5% NFDM/TBST

This blot was developed using a high-sensitivity ECL substrate, allowing for the detection of proteins in the mid-femtogram range.

ab181602 was used as a GAPDH loading control.

All lanes:

Western blot - Anti-Iba1 antibody [EPR16588] - Microglia marker – Chicken IgY (Chimeric) (ab318302) at 1/1000 dilution

Lane 1:

Mouse brain tissue lysate at 20 µg

Lane 2:

Mouse heart tissue lysate at 20 µg

Lane 3:

Mouse kidney tissue lysate at 20 µg

Lane 4:

Mouse spleen tissue lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Chicken IgY H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-chicken-igy-h-l-hrp-ab6877'>ab6877</a>) at 1/20000 dilution

Observed band size: 16 kDa

true

Exposure time: 3min