Anti-FcRgamma antibody [EPR30963-69]

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-FcRgamma antibody [EPR30963-69] (AB325712)

Immunohistochemical analysis of paraffin-embedded (A) THP-1 (human monocytic leukemia monocyte) cell pellet (B) 293T (human embryonic kidney epithelial cell) cell pellet tissue labeling with ab325712 at 1/500 (0.998 µg/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on (A) THP-1 (human monocytic leukemia monocyte) cell pellet, no staining on (B) 293T (human embryonic kidney epithelial cell) cell pellet. The primary antibody was incubated 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).

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-FcRgamma antibody [EPR30963-69] (AB325712)

Immunohistochemical analysis of paraffin-embedded Human liver cancer tissue labeling with ab325712 at 1/500 (0.998 µg/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on immune cells of human liver cancer. The primary antibody was incubated 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).

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-FcRgamma antibody [EPR30963-69] (AB325712)

Immunohistochemical analysis of paraffin-embedded Human epityphlon tissue labeling with ab325712 at 1/500 (0.998 µg/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on immune cells of human epityphlon (PMID : 36842142). The primary antibody was incubated 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).

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-FcRgamma antibody [EPR30963-69] (AB325712)

Immunohistochemical analysis of paraffin-embedded Human liver tissue labeling with ab325712 at 1/500 (0.998 µg/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on Kupffer cells of human liver (PMID : 39893200). The primary antibody was incubated 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).

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-FcRgamma antibody [EPR30963-69] (AB325712)

Immunohistochemical analysis of paraffin-embedded Human cerebrum tissue labeling with ab325712 at 1/500 (0.998 µg/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on microglia of human cerebrum (PMID : 38143775). The primary antibody was incubated 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).

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-FcRgamma antibody [EPR30963-69] (AB325712)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized THP-1 (human monocytic leukemia monocyte) 293T (human embryonic kidney epithelial cell) cells labelling with ab325712 at 1/50 (9.98 µg/ml) dilution , followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor®488) preadsorbed antibody at 1/1000 dilution (Green).

Confocal image showing cytoplasmic staining in THP-1 cell line and no staining in 293T cell line(shown in green). The counterstain was observed in magenta. Nuclear DNA was labelled with DAPI (shown in blue). Low expression : 293T. Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

ab7291 Anti-alpha Tubulin mouse monoclonal antibody was used to counterstain tubulin at 1/1000 dilution (Magenta). 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 dilution.

• IP

Lab

Immunoprecipitation - Anti-FcRgamma antibody [EPR30963-69] (AB325712)

was immunoprecipitated from 0.35 mg THP-1 (human monocytic leukemia monocyte) whole cell lysate with ab325712 at 1/30 dilution (2µg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab325712 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution.

Lane 1 : THP-1 (human monocytic leukemia monocyte) whole cell lysate
Lane 2 : ab325712 IP in THP-1 (human monocytic leukemia monocyte) whole cell lysate
Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab325712 in THP-1 whole cell lysate

Blocking and dilution buffer and concentration : 5% NFDM/TBST.

Exposure time : 24 seconds.

All lanes:

Immunoprecipitation - Anti-FcRgamma antibody [EPR30963-69] (ab325712) at 1/1000 dilution

Lane 1:

THP-1 (human monocytic leukemia monocyte) whole cell lysate at 5 µg

Lane 2:

ab325712 at 1/30 IP in THP-1 (human monocytic leukemia monocyte) whole cell lysate

Lane 3:

Rabbit monoclonal IgG (<a href='/en-us/products/primary-antibodies/rabbit-igg-monoclonal-epr25a-isotype-control-ab172730'>ab172730</a>) instead of ab325712 in THP-1 whole cell lysate

Secondary

All lanes:

Immunoprecipitation - VeriBlot for IP Detection Reagent (HRP) (<a href='/en-us/products/reagents/veriblot-for-ip-detection-reagent-hrp-ab131366'>ab131366</a>) at 1/5000 dilution

Observed band size: 10 kDa

false

Exposure time: 24s

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-FcRgamma antibody [EPR30963-69] (AB325712)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized RAW 264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage) Neuro-2a (mouse neuroblastoma neuroblast) cells labelling with ab325712 at 1/50 (9.98 µg/ml) dilution , followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor®488) preadsorbed antibody at 1/1000 dilution (Green).

Confocal image showing cytoplasmic staining in RAW 264.7 cell line and no staining in Neuro-2a cell line(shown in green). The counterstain was observed in magenta. Nuclear DNA was labelled with DAPI (shown in blue). Low expression : Neuro-2a. Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

ab7291 Anti-alpha Tubulin mouse monoclonal antibody was used to counterstain tubulin at 1/1000 dilution (Magenta). 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 dilution.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-FcRgamma antibody [EPR30963-69] (AB325712)

Immunohistochemical analysis of paraffin-embedded Rat cerebrum tissue labeling with ab325712 at 1/500 (0.998 µg/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on microglia of rat cerebrum. The primary antibody was incubated 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).

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-FcRgamma antibody [EPR30963-69] (AB325712)

Immunohistochemical analysis of paraffin-embedded Rat spleen tissue labeling with ab325712 at 1/500 (0.998 µg/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on rat spleen. The primary antibody was incubated 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).

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-FcRgamma antibody [EPR30963-69] (AB325712)

Immunohistochemical analysis of paraffin-embedded Mouse cerebrum tissue labeling with ab325712 at 1/500 (0.998 µg/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on microglia of mouse cerebrum. The primary antibody was incubated 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).

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-FcRgamma antibody [EPR30963-69] (AB325712)

Immunohistochemical analysis of paraffin-embedded Mouse liver cancer tissue labeling with ab325712 at 1/500 (0.998 µg/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on immune cells of mouse liver cancer. The primary antibody was incubated 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).

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-FcRgamma antibody [EPR30963-69] (AB325712)

Immunohistochemical analysis of paraffin-embedded Rat liver tissue labeling with ab325712 at 1/500 (0.998 µg/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on Kupffer cells of rat liver. The primary antibody was incubated 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).

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-FcRgamma antibody [EPR30963-69] (AB325712)

Immunohistochemical analysis of paraffin-embedded Mouse liver tissue labeling with ab325712 at 1/500 (0.998 µg/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on Kupffer cells of mouse liver. The primary antibody was incubated 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).

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-FcRgamma antibody [EPR30963-69] (AB325712)

Immunohistochemical analysis of paraffin-embedded Mouse spleen tissue labeling with ab325712 at 1/500 (0.998 µg/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on mouse spleen. The primary antibody was incubated 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).

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-FcRgamma antibody [EPR30963-69] (AB325712)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized Rat spleenocytes cells labelling with ab325712 at 1/50 (9.98 µg/ml) dilution , followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor®488) preadsorbed antibody at 1/1000 dilution (Green).

Confocal image showing cytoplasmic staining in subsets of Rat spleenocytes (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).

ab7291 Anti-alpha Tubulin mouse monoclonal antibody was used to counterstain tubulin at 1/1000 dilution (Magenta). 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 dilution.

• IP

Lab

Immunoprecipitation - Anti-FcRgamma antibody [EPR30963-69] (AB325712)

was immunoprecipitated from 0.35 mg RAW 264.7 (mouse abelson murine leukemia virus-induced tumor macrophage) whole cell lysate with ab325712 at 1/30 dilution (2µg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab325712 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution.

Lane 1 : RAW 264.7 (mouse abelson murine leukemia virus-induced tumor macrophage) whole cell lysate
Lane 2 : ab325712 IP in RAW 264.7 (mouse abelson murine leukemia virus-induced tumor macrophage) whole cell lysate
Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab325712 in RAW 264.7 whole cell lysate

Blocking and dilution buffer and concentration : 5% NFDM/TBST.

Exposure time : 24 seconds.

All lanes:

Immunoprecipitation - Anti-FcRgamma antibody [EPR30963-69] (ab325712) at 1/1000 dilution

Lane 1:

RAW 264.7 (mouse abelson murine leukemia virus-induced tumor macrophage) whole cell lysate at 5 µg

Lane 2:

ab325712 at 1/30 IP in RAW 264.7 (mouse abelson murine leukemia virus-induced tumor macrophage) whole cell lysate

Lane 3:

Rabbit monoclonal IgG (<a href='/en-us/products/primary-antibodies/rabbit-igg-monoclonal-epr25a-isotype-control-ab172730'>ab172730</a>) instead of ab325712 in RAW 264.7 whole cell lysate

Secondary

All lanes:

Immunoprecipitation - VeriBlot for IP Detection Reagent (HRP) (<a href='/en-us/products/reagents/veriblot-for-ip-detection-reagent-hrp-ab131366'>ab131366</a>) at 1/5000 dilution

Observed band size: 10 kDa

false

Exposure time: 24s

• IP

Lab

Immunoprecipitation - Anti-FcRgamma antibody [EPR30963-69] (AB325712)

was immunoprecipitated from 0.35 mg NR8383 (rat lung macrophage (alveolar)) whole cell lysate with ab325712 at 1/30 dilution (2µg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab325712 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution.

Lane 1 : NR8383 (rat lung macrophage (alveolar)) whole cell lysate
Lane 2 : ab325712 IP in NR8383 (rat lung macrophage (alveolar)) whole cell lysate
Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab325712 in NR8383 whole cell lysate

Blocking and dilution buffer and concentration : 5% NFDM/TBST.

Exposure time : 8 seconds.

All lanes:

Immunoprecipitation - Anti-FcRgamma antibody [EPR30963-69] (ab325712) at 1/1000 dilution

Lane 1:

NR8383 (rat lung macrophage (alveolar)) whole cell lysate at 5 µg

Lane 2:

ab325712 at 1/30 IP in NR8383 (rat lung macrophage (alveolar)) whole cell lysate

Lane 3:

Rabbit monoclonal IgG (<a href='/en-us/products/primary-antibodies/rabbit-igg-monoclonal-epr25a-isotype-control-ab172730'>ab172730</a>) instead of ab325712 in NR8383 whole cell lysate

Secondary

All lanes:

Immunoprecipitation - VeriBlot for IP Detection Reagent (HRP) (<a href='/en-us/products/reagents/veriblot-for-ip-detection-reagent-hrp-ab131366'>ab131366</a>) at 1/5000 dilution

Observed band size: 10 kDa

false

Exposure time: 8s

• WB

Lab

Western blot - Anti-FcRgamma antibody [EPR30963-69] (AB325712)

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

Lanes 1 was incubated with Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 and Lanes 2-3 were incubated with Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1/20000.

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) (1 : 200000) (36KDa);.

Exposure time : Lane 1 : 10 seconds, lane 2 : 26 seconds, lane 3 : 10 seconds

All lanes:

Western blot - Anti-FcRgamma antibody [EPR30963-69] (ab325712) at 1/1000 dilution

Lane 1:

Human spleen tissue lysate at 40 µg

Lane 2:

Mouse spleen tissue lysate at 40 µg

Lane 3:

Rat spleen tissue lysate at 40 µg

Secondary

Lane 1:

Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution

Lanes 2 - 3:

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: 10 kDa,36 kDa

false

• WB

Lab

Western blot - Anti-FcRgamma antibody [EPR30963-69] (AB325712)

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

Low expression : 293T, SK-OV-3, Neuro-2a

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) (1 : 200000) (36KDa);.

All lanes:

Western blot - Anti-FcRgamma antibody [EPR30963-69] (ab325712) at 1/1000 dilution

Lane 1:

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

Lane 2:

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

Lane 3:

SK-OV-3 (human ovarian cancer epithelial cell) whole cell lysate at 20 µg

Lane 4:

RAW 264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage) whole cell lysate at 20 µg

Lane 5:

Neuro-2a (mouse neuroblastoma neuroblast)whole cell lysate at 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: 10 kDa,36 kDa

false

Exposure time: 15s