Anti-p40-phox antibody [EP2142Y] - BSA and Azide free

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-p40-phox antibody [EP2142Y] - BSA and Azide free (AB247448)

This data was developed using ab76158, the same antibody clone in a different buffer formulation.

ab76158 at 1/100-1/250 dilution staining p40-phox in human spleen by Immunohistochemistry, Paraffin-embedded tissue.

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-p40-phox antibody [EP2142Y] - BSA and Azide free (AB247448)

This data was developed using ab76158, the same antibody clone in a different buffer formulation.

ICC/IF image of ab76158 stained MCF7 cells. The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab76158, 1/25 dilution) overnight at +4°C. The secondary antibody (green) was ab96899, DyLight® 488 goat anti-rabbit IgG (H+L) used at a 1/250 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43μM.

• Flow Cyt (Intra)

Unknown

Flow Cytometry (Intracellular) - Anti-p40-phox antibody [EP2142Y] - BSA and Azide free (AB247448)

This data was developed using ab76158, the same antibody clone in a different buffer formulation.Overlay histogram showing K562 cells stained with ab76158 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab76158, 1/100 dilution) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-rabbit IgG (H+L) (ab96899) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (1μg/1x10⁶ cells) used under the same conditions. Acquisition of >5,000 events was performed.

• IP

Lab

Immunoprecipitation - Anti-p40-phox antibody [EP2142Y] - BSA and Azide free (AB247448)

This data was developed using ab76158, the same antibody clone in a different buffer formulation.

p40-phox was immunoprecipitated from 0.35 mg HL-60 (Human acute promyelocytic leukemia promyeloblast) whole cell lysate 10 μg with ab76158 at 1/50 dilution (2μg). VeriBlot for IP Detection Reagent (HRP) (ab131366) was used at 1/5000 dilution.

Lane 1 : HL-60 (Human acute promyelocytic leukemia promyeloblast) whole cell lysate 10 μg

Lane 2 : ab76158 IP in HL-60 whole cell lysate

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab76158 in HL-60 whole cell lysate

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

All lanes:

Immunoprecipitation - Anti-p40-phox antibody [EP2142Y] (<a href='/en-us/products/primary-antibodies/p40-phox-antibody-ep2142y-ab76158'>ab76158</a>)

Predicted band size: 39 kDa

Observed band size: 39 kDa

false

• WB

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Western blot - Anti-p40-phox antibody [EP2142Y] - BSA and Azide free (AB247448)

This data was developed using ab76158, the same antibody clone in a different buffer formulation.

All lanes:

Western blot - Anti-p40-phox antibody [EP2142Y] (<a href='/en-us/products/primary-antibodies/p40-phox-antibody-ep2142y-ab76158'>ab76158</a>) at 1/50000 dilution

All lanes:

HL60 cell lysate at 10 µg

Secondary

All lanes:

Goat anti-rabbit HRP conjugated at 1/2000 dilution

Predicted band size: 39 kDa

Observed band size: 39 kDa

false