Anti-F4/80 antibody [EPR26545-166] (BSA and Azide free)

• mIHC

Lab

Multiplex immunohistochemistry - Anti-F4/80 antibody [EPR26545-166] (BSA and Azide free) (AB300422)

This data was developed using the same antibody clone in a different buffer formulation ( ab300421).

Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded Mouse thymus tissue staining Neutrophil Elastase with ab310335 at a 1/500 ( 1.076 µg/ml) dilution, F4/80 with ab300421 at a 1/5000 ( 0.106 µg/ml) dilution and NCR1 with ab233558 at a 1/500 ( 1.114 µg/ml) dilution followed by a ready to use secondary antibody Opal Polymer HRP Ms + Rb.

Panel A : merged staining of anti-Neutrophil Elastase (magenta; Opal™690), anti-CD19 (green; Opal™520) and anti-CD3 (gray; Opal™690) on mouse spleen.Panel B : anti-Neutrophil Elastase staining neutrophils in mouse spleen.
Panel C : anti-CD19 staining B lymphocytes in mouse spleen.
Panel D : anti-CD3 staining T lymphocytes in mouse spleen.
Nuclear DNA was labeled with DAPI (shown in blue).

The section was incubated in three rounds of staining : in the order of ab310335, ab245235 and ab237721 for 30 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system.

The immunostaining was performed on a Leica Biosystems BOND^® RX instrument with an Opal™ 4-color kit. Image acquisition was performed with Leica SP8 confocal microscope.

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-F4/80 antibody [EPR26545-166] (BSA and Azide free) (AB300422)

This data was developed using ab300421, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Rat liver tissue labeling F4/80 with ab300421 at 1/5000 (0.1 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) was used. Positive staining on the Kupffer cells of rat liver. The section was incubated with ab300421 for 30 mins at room temperature. 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) was used. 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-F4/80 antibody [EPR26545-166] (BSA and Azide free) (AB300422)

This data was developed using ab300421, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Mouse spleen tissue labeling F4/80 with ab300421 at 1/5000 (0.1 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) was used. Positive staining on the macrophages of mouse spleen (PMID : 19820169). The section was incubated with ab300421 for 30 mins at room temperature. 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) was used. 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-F4/80 antibody [EPR26545-166] (BSA and Azide free) (AB300422)

This data was developed using ab300421, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Mouse liver tissue labeling F4/80 with ab300421 at 1/5000 (0.1 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) was used. Positive staining on the Kupffer cells of mouse liver (PMID : 26473015). The section was incubated with ab300421 for 30 mins at room temperature.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) was used. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-F4/80 antibody [EPR26545-166] (BSA and Azide free) (AB300422)

This data was developed using ab300421, the same antibody clone in a different buffer formulation.Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized RAW 264.7 (mouse abelson murine leukemia virus-induced tumor macrophage) cells labelling F4/80 with ab300421 at 1/50 dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 2ug/mL dilution (Green). Confocal image showing membranous and cytoplasmic staining in RAW264.7 cell lineNegative control : NIH/3T3 (PMID : 8550607) is observed. ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 2.5ug/ml dilution (Red). The Nuclear counterstain was DAPI (Blue). Secondary antibody only control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2ug/mL dilution.

• IHC-Fr

Supplier Data

Immunohistochemistry (Frozen sections) - Anti-F4/80 antibody [EPR26545-166] (BSA and Azide free) (AB300422)

This data was developed using ab300421, the same antibody clone in a different buffer formulation. Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Mouse spleen (fresh) tissue labeling F4/80 with ab300421 at 1/500 dilution followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2 ug/mL dilution (Green). Positive staining on the macrophages of mouse spleen is observed. The nuclear counterstain was DAPI (Blue). Secondary antibody control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2 ug/mL dilution.

• mIHC

Lab

Multiplex immunohistochemistry - Anti-F4/80 antibody [EPR26545-166] (BSA and Azide free) (AB300422)

This data was developed using ab300421, the same antibody clone in a different buffer formulation.

Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded Mouse spleen staining of F4/80 with ab300421 at a 1/5000 (0.1 µg/ml) dilution, CD19 with ab245235 at 1/1000 (0.444 µg/ml) dilution and CD3 with ab237721 at 1/2000 ( 0.264 µg/ml) dilution.

Panel A : merged staining of anti-F4/80 (magenta; Opal™690), anti-CD19 (green; Opal™520) and anti-CD3 (gray; Opal™570) on mouse spleen.
Panel B : anti-F4/80 staining macrophages in mouse spleen.
Panel C : anti-CD19 staining B lymphocytes in mouse spleen.
Panel D : anti-CD3 staining T lymphocytes in mouse spleen.
Nuclear DNA was labelled with DAPI (shown in blue).

The section was incubated in three rounds of staining : in the order of ab300421, ab245235 and ab237721 for 30 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system.

The immunostaining was performed on a Leica Biosystems BOND^® RX instrument with an Opal™ 4-color kit. Image acquisition was performed with Leica SP8 confocal microscope.

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0 epitope retrieval solution2) for 20 mins.

• mIHC

Lab

Multiplex immunohistochemistry - Anti-F4/80 antibody [EPR26545-166] (BSA and Azide free) (AB300422)

This data was developed using the same antibody clone in a different buffer formulation ( ab300421).

Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded Mouse spleen tissue staining Neutrophil Elastase with ab310335 at a 1/500 ( 1.076 µg/ml) dilution, F4/80 with ab300421 at a 1/5000 ( 0.106 µg/ml) dilution and NCR1 with ab233558 at a 1/500 ( 1.114 µg/ml) dilution followed by a ready to use secondary antibody Opal Polymer HRP Ms + Rb.

Panel A : merged staining of anti-Neutrophil Elastase (magenta; Opal™690), anti-CD19 (green; Opal™520) and anti-CD3 (gray; Opal™690) on mouse spleen.Panel B : anti-Neutrophil Elastase staining neutrophils in mouse spleen.
Panel C : anti-CD19 staining B lymphocytes in mouse spleen.
Panel D : anti-CD3 staining T lymphocytes in mouse spleen.
Nuclear DNA was labeled with DAPI (shown in blue).

The section was incubated in three rounds of staining : in the order of ab310335, ab245235 and ab237721 for 30 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system.

The immunostaining was performed on a Leica Biosystems BOND^® RX instrument with an Opal™ 4-color kit. Image acquisition was performed with Leica SP8 confocal microscope.

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-F4/80 antibody [EPR26545-166] (BSA and Azide free) (AB300422)

This data was developed using ab300421, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Rat spleen tissue labeling F4/80 with ab300421 at 1/5000 (0.1 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) was used. Positive staining on the macrophages of rat spleen. The section was incubated with ab300421 for 30 mins at room temperature. 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) was used. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

• mIHC

Lab

Multiplex immunohistochemistry - Anti-F4/80 antibody [EPR26545-166] (BSA and Azide free) (AB300422)

This data was developed using ab300421, the same antibody clone in a different buffer formulation.

Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded rat lung staining of Surfactant Protein A/PSAP with ab322404 at a 1/5000 (0.092 μg/ml) dilution, F4/80 with ab300421 at 1/5000 (0.070 μg/ml) dilution, and CD73 with ab314327 at 1/500 (0.98 μg/ml), followed by a ready to use secondary antibody Opal Polymer HRP Ms + Rb.

Panel A : merged staining of anti-Surfactant Protein A/PSAP (green; Opal™ 520), anti-F4/80 (grey; Opal™ 570) and anti-CD73 (magenta; Opal™ 690) on rat lung.
Panel B : anti-Surfactant Protein A/PSAP staining Clara cells and alveolar type II cells in rat lung.
Panel C : anti-F4/80 staining macrophages in rat lung.
Panel D : anti-CD73 staining epithelium in rat lung.
Nuclear DNA was labeled with DAPI (shown in blue).

The section was incubated in three rounds of staining : in the order of ab322404, ab300421 and ab314327 for 30 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system..

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

The immunostaining was performed on a Leica Biosystems BOND® RX instrument with an Opal™ 4-color kit. Image acquisition was performed with Leica SP8 confocal microscope.

• mIHC

Lab

Multiplex immunohistochemistry - Anti-F4/80 antibody [EPR26545-166] (BSA and Azide free) (AB300422)

This data was developed using ab300421, the same antibody clone in a different buffer formulation.

Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded mouse lung staining of Surfactant Protein A/PSAP with ab322404 at a 1/5000 (0.092 μg/ml) dilution, F4/80 with ab300421 at 1/5000 (0.070 μg/ml) dilution, and CD73 with ab314327 at 1/500 (0.98 μg/ml), followed by a ready to use secondary antibody Opal Polymer HRP Ms + Rb.

Panel A : merged staining of anti-Surfactant Protein A/PSAP (green; Opal™ 520), anti-F4/80 (grey; Opal™ 570) and anti-CD73 (magenta; Opal™ 690) on mouse lung.
Panel B : anti-Surfactant Protein A/PSAP staining Clara cells and alveolar type II cells in mouse lung.
Panel C : anti-F4/80 staining macrophages in mouse lung.
Panel D : anti-CD73 staining epithelium in mouse lung.
Nuclear DNA was labeled with DAPI (shown in blue).

The section was incubated in three rounds of staining : in the order of ab322404, ab300421 and ab314327 for 30 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system..

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

The immunostaining was performed on a Leica Biosystems BOND® RX instrument with an Opal™ 4-color kit. Image acquisition was performed with Leica SP8 confocal microscope.

• mIHC

Lab

Multiplex immunohistochemistry - Anti-F4/80 antibody [EPR26545-166] (BSA and Azide free) (AB300422)

This data was developed using ab300421, the same antibody clone in a different buffer formulation.

Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded Mouse bone marrow staining of F4/80 with ab300421 at a 1/5000 (0.1 µg/ml) dilution, CD19 with ab245235 at 1/1000 (0.444 µg/ml) dilution and CD3 with ab237721 at 1/2000 ( 0.264 µg/ml) dilution.

Panel A : merged staining of anti-F4/80 (magenta; Opal™690), anti-CD19 (green; Opal™520) and anti-CD3 (gray; Opal™570) on mouse bone marrow.
Panel B : anti-F4/80 staining macrophages in mouse bone marrow.
Panel C : anti-CD19 staining B lymphocytes in mouse bone marrow.
Panel D : anti-CD3 staining T lymphocytes in mouse bone marrow.
Nuclear DNA was labelled with DAPI (shown in blue).

The section was incubated in three rounds of staining : in the order of ab300421, ab245235 and ab237721 for 30 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system.

The immunostaining was performed on a Leica Biosystems BOND^® RX instrument with an Opal™ 4-color kit. Image acquisition was performed with Leica SP8 confocal microscope.

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0 epitope retrieval solution2) for 20 mins.

• mIHC

Lab

Multiplex immunohistochemistry - Anti-F4/80 antibody [EPR26545-166] (BSA and Azide free) (AB300422)

This data was developed using ab300421, the same antibody clone in a different buffer formulation.

Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded Mouse thymus staining of F4/80 with ab300421 at a 1/5000 (0.1 µg/ml) dilution, CD19 with ab245235 at 1/1000 (0.444 µg/ml) dilution and CD3 with ab237721 at 1/2000 ( 0.264 µg/ml) dilution.

Panel A : merged staining of anti-F4/80 (magenta; Opal™690), anti-CD19 (green; Opal™520) and anti-CD3 (gray; Opal™570) on mouse thymus.
Panel B : anti-F4/80 staining macrophages in mouse thymus.
Panel C : anti-CD19 staining B lymphocytes in mouse thymus.
Panel D : anti-CD3 staining T lymphocytes in mouse thymus.
Nuclear DNA was labelled with DAPI (shown in blue).

The section was incubated in three rounds of staining : in the order of ab300421, ab245235 and ab237721 for 30 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system.

The immunostaining was performed on a Leica Biosystems BOND^® RX instrument with an Opal™ 4-color kit. Image acquisition was performed with Leica SP8 confocal microscope.

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0 epitope retrieval solution2) for 20 mins.

• mIHC

Lab

Multiplex immunohistochemistry - Anti-F4/80 antibody [EPR26545-166] (BSA and Azide free) (AB300422)

This data was developed using ab300421, the same antibody clone in a different buffer formulation.

Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded Mouse lymph node staining of F4/80 with ab300421 at a 1/5000 (0.1 µg/ml) dilution, CD19 with ab245235 at 1/1000 (0.444 µg/ml) dilution and CD3 with ab237721 at 1/2000 ( 0.264 µg/ml) dilution.

Panel A : merged staining of anti-F4/80 (magenta; Opal™690), anti-CD19 (green; Opal™520) and anti-CD3 (gray; Opal™570) on mouse lymph node.
Panel B : anti-F4/80 staining macrophages in mouse lymph node.
Panel C : anti-CD19 staining B lymphocytes in mouse lymph node.
Panel D : anti-CD3 staining T lymphocytes in mouse lymph node.
Nuclear DNA was labelled with DAPI (shown in blue).

The section was incubated in three rounds of staining : in the order of ab300421, ab245235 and ab237721 for 30 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system.

The immunostaining was performed on a Leica Biosystems BOND^® RX instrument with an Opal™ 4-color kit. Image acquisition was performed with Leica SP8 confocal microscope.

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0 epitope retrieval solution2) for 20 mins.

• mIHC

Lab

Multiplex immunohistochemistry - Anti-F4/80 antibody [EPR26545-166] (BSA and Azide free) (AB300422)

This data was developed using the same antibody clone in a different buffer formulation ( ab300421).

Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded Mouse lymph node tissue staining Neutrophil Elastase with ab310335 at a 1/500 ( 1.076 µg/ml) dilution, F4/80 with ab300421 at a 1/5000 ( 0.106 µg/ml) dilution and NCR1 with ab233558 at a 1/500 ( 1.114 µg/ml) dilution followed by a ready to use secondary antibody Opal Polymer HRP Ms + Rb.

Panel A : merged staining of anti-Neutrophil Elastase (magenta; Opal™690), anti-CD19 (green; Opal™520) and anti-CD3 (gray; Opal™690) on mouse spleen.Panel B : anti-Neutrophil Elastase staining neutrophils in mouse spleen.
Panel C : anti-CD19 staining B lymphocytes in mouse spleen.
Panel D : anti-CD3 staining T lymphocytes in mouse spleen.
Nuclear DNA was labeled with DAPI (shown in blue).

The section was incubated in three rounds of staining : in the order of ab310335, ab245235 and ab237721 for 30 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system.

The immunostaining was performed on a Leica Biosystems BOND^® RX instrument with an Opal™ 4-color kit. Image acquisition was performed with Leica SP8 confocal microscope.

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins

• WB

Supplier Data

Western blot - Anti-F4/80 antibody [EPR26545-166] (BSA and Azide free) (AB300422)

This data was developed using ab300421, the same antibody clone in a different buffer formulation. Blocking Buffer and concentration : 5% NFDM/TBST

All lanes:

Western blot - Anti-F4/80 antibody [EPR26545-166] (<a href='/en-us/products/primary-antibodies/f4-80-antibody-epr26545-166-ab300421'>ab300421</a>) at 1/1000 dilution

All lanes:

Mouse spleen tissue lysate 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Observed band size: 160 kDa

false

Exposure time: 26s

• WB

Supplier Data

Western blot - Anti-F4/80 antibody [EPR26545-166] (BSA and Azide free) (AB300422)

This data was developed using ab300421, the same antibody clone in a different buffer formulation. Blocking Buffer and concentration : 5% NFDM/TBST

All lanes:

Western blot - Anti-F4/80 antibody [EPR26545-166] (<a href='/en-us/products/primary-antibodies/f4-80-antibody-epr26545-166-ab300421'>ab300421</a>) at 1/1000 dilution

Lane 1:

J774A.1 (mouse reticum cell sarcoma macrophage) whole cell lysate 20

Lane 2:

NIH/3T3 (mouse embryonic fibroblast) whole cell lysate

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Observed band size: 90 kDa,160 kDa

false

Exposure time: 15s

• WB

Supplier Data

Western blot - Anti-F4/80 antibody [EPR26545-166] (BSA and Azide free) (AB300422)

This data was developed using ab300421, the same antibody clone in a different buffer formulation. Blocking Buffer and concentration : 5% NFDM/TBST

All lanes:

Western blot - Anti-F4/80 antibody [EPR26545-166] (<a href='/en-us/products/primary-antibodies/f4-80-antibody-epr26545-166-ab300421'>ab300421</a>) at 1/1000 dilution

Lane 1:

RAW 264.7 (mouse abelson murine leukemia virus-induced tumor macrophage) whole cell lysate

Lane 2:

L-929 ( mouse connective tissue fibroblast) whole cell lysate

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Observed band size: 160 kDa

false

Exposure time: 15s