Anti-NCF1/p47-phox antibody [EPR27205-231]

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-NCF1/p47-phox antibody [EPR27205-231] (AB308256)

Immunofluorescent analysis of 80% Methanol-fixed, 0.1% Triton X-100 permeabilized RAW 264.7 (mouse abelson murine leukemia virus-induced tumor macrophage) cells labelling NCF1/p47-phox with ab308256 at 1/250 dilution (2.004 ug/ml) followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 dilution (2 ug/ml) (Green). Confocal image showing cytoplasmic and weak nuclear staining in RAW 264.7 cell line, no staining was observed in HL-1 cell line. Low expression control : HL-1. Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8). ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 dilution (2.5 ug/ml) (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 dilution (2 ug/ml).

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-NCF1/p47-phox antibody [EPR27205-231] (AB308256)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized HL-1 (mouse atrial muscle cell) (Left panel) compared to RAW 264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage) (Right panel) cells labelling NCF1/p47-phox with ab308256 at 1/500 dilution (0.1 ug) (Red) compared with a Rabbit monoclonal IgG (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, ab150081) at 1/5000 dilution was used as the secondary antibody. Cells were stained with rabbit IgG or ab308256

• WB

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Western blot - Anti-NCF1/p47-phox antibody [EPR27205-231] (AB308256)

Blocking and diluting buffer and concentration : 5% NFDM/TBST. Low expression : HL-1. The Lane 3 was developed using a high sensitivity ECL substrate. In lanes 1-2, the lysates were stored at -80°C prior to Western Blotting. The bands beneath the target band (32 kDa) are likely to be degradation products. In lane 3, the lysate was freshly made and used for Western Blotting immediately to minimize protein degradation. In Western blot, anti-GAPDH antibody (ab181602) loading control staining at 1/200000 dilution. Exposure time : 180 seconds.

All lanes:

Western blot - Anti-NCF1/p47-phox antibody [EPR27205-231] (ab308256) 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:

HL-1 (mouse atrial muscle cell) whole cell lysate at 20 µg

Lane 3:

RAW 264.7 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 1/100000 dilution

Predicted band size: 44 kDa

Observed band size: 45 kDa

false

Exposure time: 180s

• WB

Supplier Data

Western blot - Anti-NCF1/p47-phox antibody [EPR27205-231] (AB308256)

Blocking and diluting buffer and concentration : 5% NFDM/TBST. Low expression : skeletal muscle, brain. In Western blot, anti-GAPDH antibody (ab181602) loading control staining at 1/200000 dilution. Exposure time : 180 seconds.

All lanes:

Western blot - Anti-NCF1/p47-phox antibody [EPR27205-231] (ab308256) at 1/1000 dilution

Lane 1:

Mouse lymph node tissue lysate at 20 µg

Lane 2:

Mouse spleen tissue lysate at 20 µg

Lane 3:

Mouse skeletal muscle tissue lysate at 20 µg

Lane 4:

Mouse brain tissue lysate at 20 µg

Lane 5:

Rat spleen tissue lysate at 20 µg

Lane 6:

Rat skeletal muscle tissue 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

Predicted band size: 44 kDa

Observed band size: 45 kDa

false

Exposure time: 180s