Anti-NOX2/gp91phox antibody [EPR28415-13]

19 product images

• 

  Immunoprecipitation - Anti-NOX2/gp91phox antibody [EPR28415-13] (ab310337)

  NOX2/gp91phox was immunoprecipitated from 0.35 mg RAW 264.7 (mouse abelson murine leukemia virus-induced tumor macrophage) whole cell lysate with ab310337 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab310337 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(VeriBlot for IP Detection Reagent (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: ab310337 IP in RAW 264.7 (mouse abelson murine leukemia virus-induced tumor macrophage) whole cell lysate
  Lane 3:Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of ab310337 in RAW 264.7 whole cell lysate
  Blocking and dilution buffer and concentration: 5% NFDM/TBST.
  Exposure time: 3 seconds

  All lanes: Immunoprecipitation - Anti-NOX2/gp91phox antibody [EPR28415-13] (ab310337) at 1/30 dilution

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

  Secondary

  All lanes: Immunoprecipitation - VeriBlot for IP Detection Reagent (HRP) (VeriBlot for IP Detection Reagent (HRP) ab131366) at 1/5000 dilution

  Exposure time: 3s

• 

  Western blot - Anti-NOX2/gp91phox antibody [EPR28415-13] (ab310337)

  Blocking and diluting buffer and concentration: 5% NFDM/TBST
  Low expression: Neuro-2a (PMID: 23516464), skeletal muscle (PMID:11376945).
  
  In Western blot, anti-GAPDH antibody (Anti-GAPDH antibody [EPR16891] - Loading Control ab181602) loading control staining at 1/200, 000 dilution.
  Exposure time: Lanes 1-2: 15 seconds;
  Lanes 3-4: 59 seconds;
  Lane 5:15 seconds.

  All lanes: Western blot - Anti-NOX2/gp91phox antibody [EPR28415-13] (ab310337) at 1/1000 dilution

  Lane 1: RAW 264.7 (mouse abelson murine leukemia virus-induced tumor macrophage) whole cell lysate at 20 µg

  Lane 2: Neuro-2a (mouse neuroblastoma neuroblast) whole cell lysate at 20 µg

  Lane 3: Mouse spleen tissue lysate at 20 µg

  Lane 4: Mouse skeletal muscle tissue lysate at 20 µg

  Lane 5: NR8383 (rat alveolar macrophage) whole cell lysate at 20 µg

  Secondary

  All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/20000 dilution

  Observed band size: 65 kDa

• 

  Western blot - Anti-NOX2/gp91phox antibody [EPR28415-13] (ab310337)

  Blocking and diluting buffer and concentration: 5% NFDM/TBST
  Low expression: skeletal muscle (PMID:11376945).
  
  In Western blot, anti-GAPDH antibody (Anti-GAPDH antibody [EPR16891] - Loading Control ab181602) loading control staining at 1/200, 000 dilution.
  Exposure time: 59 seconds

  All lanes: Western blot - Anti-NOX2/gp91phox antibody [EPR28415-13] (ab310337) at 1/1000 dilution

  Lane 1: Rat spleen tissue lysate at 20 µg

  Lane 2: Rat skeletal muscle tissue lysate at 20 µg

  Secondary

  All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/20000 dilution

  Observed band size: 65 kDa

  Exposure time: 59s

• 

  Immunocytochemistry/ Immunofluorescence - Anti-NOX2/gp91phox antibody [EPR28415-13] (ab310337)

  Immunofluorescent analysis of 100% methanol-fixed, 0.1% Tween-20 permeabilized NR8383 (rat alveolar macrophage) cells labelling NOX2/gp91phox with ab310337 at 1/50 (10.32 ug/ml) dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed 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 NR8383 cell line.Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8). is observed. Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker 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 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2ug/ml dilution.

• 

  Immunocytochemistry/ Immunofluorescence - Anti-NOX2/gp91phox antibody [EPR28415-13] (ab310337)

  Immunofluorescent analysis of 100% methanol-fixed, 0.1% Tween-20 permeabilized RAW 264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage) cells labelling NOX2/gp91phox with ab310337 at 1/50 (10.32 ug/ml) dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 2ug/ml dilution (Green). Confocal image showing membranous staining in RAW 264.7 cell line, and no staining in Neuro-2a cell line.Negative control: Neuro-2a (PMID: 23516464).Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8). is observed. Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker 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 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2ug/ml dilution.

• 

  Flow Cytometry (Intracellular) - Anti-NOX2/gp91phox antibody [EPR28415-13] (ab310337)

  Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized NR8383 (rat alveolar macrophage) cells labelling NOX2/gp91phox with ab310337 at 1/50 dilution (1 ug)/Red (Red) compared with a Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat Anti-Rabbit IgG (Alexa Fluor® 488, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081) at 1/5000 dilution was used as the secondary antibody.

• 

  Flow Cytometry (Intracellular) - Anti-NOX2/gp91phox antibody [EPR28415-13] (ab310337)

  Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized Neuro-2a (mouse neuroblastoma neuroblast, Left) / RAW 264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage, Right) cells labelling NOX2/gp91phox with ab310337 at 1/50 dilution (1 ug)/Red (Red) compared with a Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat Anti-Rabbit IgG (Alexa Fluor® 488, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081) at 1/5000 dilution was used as the secondary antibody. Low expression: Neuro-2a (PMID: 23516464).

• 

  Immunohistochemistry (Frozen sections) - Anti-NOX2/gp91phox antibody [EPR28415-13] (ab310337)

  Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Mouse skeletal muscle(fresh) tissue labeling NOX2/gp91phox with ab310337 at 1/50 (10.32 ug/ml) dilution followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2 ug/mL dilution (Green). Negative control: confocal image showing no staining on mouse skeletal muscle (PMID: 11376945). The nuclear counterstain was DAPI (Blue). The section was incubated with ab310337 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). is observed. The nuclear counterstain was DAPI (Blue). Secondary antibody control: Secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbedat 1/1000 2 ug/mL dilution.

• 

  Immunohistochemistry (Frozen sections) - Anti-NOX2/gp91phox antibody [EPR28415-13] (ab310337)

  Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Mouse liver (fresh) tissue labeling NOX2/gp91phox with ab310337 at 1/50 (10.32 ug/ml) dilution followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2 ug/mL dilution (Green). Confocal image showing positive staining on mouse liver. The nuclear counterstain was DAPI (Blue). The section was incubated with ab310337 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). is observed. The nuclear counterstain was DAPI (Blue). Secondary antibody control: Secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbedat 1/1000 2 ug/mL dilution.

• 

  Immunohistochemistry (Frozen sections) - Anti-NOX2/gp91phox antibody [EPR28415-13] (ab310337)

  Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Rat skeletal muscle(fresh) tissue labeling NOX2/gp91phox with ab310337 at 1/50 (10.32 ug/ml) dilution followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2 ug/mL dilution (Green). Negative control: confocal image showing no staining on rat skeletal muscle (PMID: 11376945). The nuclear counterstain was DAPI (Blue). The section was incubated with ab310337 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). is observed. The nuclear counterstain was DAPI (Blue). Secondary antibody control: Secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbedat 1/1000 2 ug/mL dilution.

• 

  Immunohistochemistry (Frozen sections) - Anti-NOX2/gp91phox antibody [EPR28415-13] (ab310337)

  Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Rat liver (fresh) tissue labeling NOX2/gp91phox with ab310337 at 1/50 (10.32 ug/ml) dilution followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2 ug/mL dilution (Green). Confocal image showing positive staining on rat liver. The nuclear counterstain was DAPI (Blue). The section was incubated with ab310337 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). is observed. The nuclear counterstain was DAPI (Blue). Secondary antibody control: Secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbedat 1/1000 2 ug/mL dilution.

• 

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NOX2/gp91phox antibody [EPR28415-13] (ab310337)

  Immunohistochemical analysis of paraffin-embedded (A) HEK-293T (human tissue labeling NOX2/gp91phox with ab310337 at 1/1000 (0.516 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on (A) HEK-293T transfected with a CYBB expression vector containing a his tag. No staining on (B) HEK-293T transfected with empty vector containing a his tag. The section was incubated with ab310337 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

• 

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NOX2/gp91phox antibody [EPR28415-13] (ab310337)

  Immunohistochemical analysis of paraffin-embedded Rat skeletal muscle tissue labeling NOX2/gp91phox with ab310337 at 1/1000 (0.516 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Negative control: No staining on rat skeletal muscle.The section was incubated with ab310337 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

• 

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NOX2/gp91phox antibody [EPR28415-13] (ab310337)

  Immunohistochemical analysis of paraffin-embedded Mouse skeletal muscl tissue labeling NOX2/gp91phox with ab310337 at 1/1000 (0.516 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Negative control: No staining on mouse skeletal muscle.The section was incubated with ab310337 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

• 

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NOX2/gp91phox antibody [EPR28415-13] (ab310337)

  Immunohistochemical analysis of paraffin-embedded Human skeletal muscl tissue labeling NOX2/gp91phox with ab310337 at 1/1000 (0.516 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Negative control: No staining on human skeletal muscle.The section was incubated with ab310337 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

• 

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NOX2/gp91phox antibody [EPR28415-13] (ab310337)

  Immunohistochemical analysis of paraffin-embedded Rat liver tissue labeling NOX2/gp91phox with ab310337 at 1/1000 (0.516 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on kupffer cells in rat liver.The section was incubated with ab310337 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

• 

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NOX2/gp91phox antibody [EPR28415-13] (ab310337)

  Immunohistochemical analysis of paraffin-embedded Mouse liver tissue labeling NOX2/gp91phox with ab310337 at 1/1000 (0.516 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on kupffer cells in mouse liver.The section was incubated with ab310337 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

• 

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NOX2/gp91phox antibody [EPR28415-13] (ab310337)

  Immunohistochemical analysis of paraffin-embedded Human spleen tissue labeling NOX2/gp91phox with ab310337 at 1/1000 (0.516 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on human spleen.The section was incubated with ab310337 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

• 

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NOX2/gp91phox antibody [EPR28415-13] (ab310337)

  Immunohistochemical analysis of paraffin-embedded Human liver tissue labeling NOX2/gp91phox with ab310337 at 1/1000 (0.516 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on kupffer cells in human liver.The section was incubated with ab310337 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