Neutrophil cytosol factor 1, also known as p47phox, is a protein that in humans is encoded by the NCF1 gene. The protein encoded by this gene is a 47 kDa cytosolic subunit of neutrophil NADPH oxidase. This oxidase is a multicomponent enzyme that is activated to produce superoxide anion. Mutations in this gene have been associated with chronic granulomatous disease. p47 is vital to the activation of NADPH oxidase. P47 becomes heavily phosphorylated. Anti-p47-phox - http://www.stjohnslabs.com/p47-phox-antibody-p-93740?filter_name=stj94885 Join our Antibody Validation Project - http://www.stjohnslabs.com/services/antibody-validation

1. Result
“In slice culture lysates, there were target bands detected by STJ91464 Anti-p47-phox antibody.“- R. Ferreira , IBMC
Portugal.
Gel electrophoresis information：10% polyacrylamide gel, constant voltage 100 V, 60 min.
Transfer information：0.45 nm PVDF membrane, 1 piece of gel (semi-dry transfer), constant voltage 25 V 27 min.
Lane No. Antigen
Loading
amount
Primary anti-
body
Primary anti-
body dilution
ratio
Secondary
antibody dilution
ratio
Target band KD
Visualiza-
tion time
1
Slice cultures
lysate
30ug
STJ94885 Anti-
p47-phox Anti-
body
1:1000
~44KD 10S
1:10000
2
Slice cultures
lysate
~44KD 10S
Cell Line:
Murine brain tissue (obtained
from organotypic slice cul-
tures)
Method of validation: Western Blot
Primary Antibody: STJ94885 Anti-p47-phox Anti-
Secondary Antibody HRP goat anti-rabbit
Reference Antibody
GAPDH (provided by Milli-
pore)
Dilution ratio: 1:1000
Antibody Customer Review:
STJ94885 Anti-p47-phox Antibody
Antibody Specificity:
Antibody Rating:
Protocol
Treatment of materials: Organotypic slice cultures were subjected to hypoxia for 1 hour, followed by treatments
E.g. Gel electrophoresis: 10% polyacrylamide gel, constant voltage 100 V, 60 min.
Transfer: 0.45 nm PVDF membrane, 1 piece of gel (semi-dry transfer), constant voltage 25 V 27 min.
Blocking: 1X TBST with 5% bovine serum albumin (BSA) for 1 hr.
Primary antibody probing: primary antibody was added to the solution of 1X TBST with 5% BSA, 4˚C for overnight.
Membrane wash: 1X TBST wash for 3 times.
Secondary antibody probing: secondary antibody was added to the solution of 1X TBST with 5% BSA with 1:10000 dilu-
tion for 1 hr.
Membrane wash: 1X TBST wash for 3 times.
Visualization: ECL visualization for 5 min (the visualization time was determined by actual result. It was ensured that
the visualization time was identical for the same antigen).
Figure: Lane1: without hypoxia; Lane 2: subjected to
hypoxia