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Invitrogen

p47phox Polyclonal Antibody

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3 Published Figures

View all (21) p47phox antibodies

Questions & Answers

Questions & Answers

Application

p47phox Antibody in Immunocytochemistry (ICC/IF)

FIGURE: 1 / 11

p47phox Antibody (PA1-9073) in ICC/IF

Immunofluorescence analysis of p47phox was performed using 70% confluent log phase Raji cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 15 minutes, and blocked with 2% BSA for 1 hour at room temperature. The cells were labeled with p47phox Goat Polyclonal Antibody (Product # PA1-9073) at 5 µg/mL in 0.1% BSA, incubated at 4 degree Celsius overnight and then labeled with Rabbit anti-Goat IgG (H+L) Cross-Adsorbed Secondary Antibody, Alexa Fluor 488 (Product # A-11078) at a dilutio... View More

p47phox Antibody in Immunocytochemistry (ICC/IF)

p47phox Antibody in Immunocytochemistry (ICC/IF)

p47phox Antibody in Immunohistochemistry (Paraffin) (IHC (P))

p47phox Antibody in Western Blot (WB)

p47phox Antibody in Western Blot (WB)

p47phox Antibody in Western Blot (WB)

p47phox Antibody in Flow Cytometry (Flow)

p47phox Antibody in Western Blot (WB)

p47phox Antibody in Western Blot (WB)

p47phox Antibody in Western Blot (WB)

p47phox Antibody

Product Image Gallery

p47phox Antibody in Immunocytochemistry (ICC/IF) — Immunofluorescence analysis of p47phox was performed using 70% confluent log phase Raji cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 15 minutes, and blocked with 2% BSA for 1 hour at room temperature. The cells were labeled with p47phox Goat Polyclonal Antibody (Product # PA1-9073) at 5 µg/mL in 0.1% BSA, incubated at 4 degree Celsius overnight and then labeled with Rabbit anti-Goat IgG (H+L) Cross-Adsorbed Secondary Antibody, Alexa Fluor 488 (Product # A-11078) at a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with ProLong™ Diamond Antifade Mountant with DAPI (Product # P36962). F-actin (Panel c: red) was stained with Rhodamine Phalloidin (Product # R415). Panel d represents the merged image showing cytoplasmic and membrane localization. Panel e represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.

p47phox Antibody in Immunocytochemistry (ICC/IF) — Immunocytochemical analysis of p47phox in HeLa cells using a p47phox polyclonal antibody (Product #PA1-9073). HeLa cells were permeabilized with 0.15% Triton. Cells were incubated with 10 µg/mL of primary antibody for one hour followed by an Alexa Fluor 488 secondary antibody at a concentration of 2 µg/mL. Cytoplasmic staining can be seen as shown above. The nuclear stain is DAPI (blue). Negative control: Unimmunized goat IgG (10 µg/mL) followed by an Alexa Fluor 488 secondary antibody (2 µg/mL).

p47phox Antibody in Immunohistochemistry (Paraffin) (IHC (P)) — Immunohistochemistry analysis of p47phox in human colon, neutrophils. Samples were incubated with p47phox polyclonal antibody (Product # PA1-9073) using a dilution of 5 µg/mL. Formalin-fixed, paraffin-embedded tissue after heat-induced antigen retrieval.

p47phox Antibody in Western Blot (WB) — Western blot was performed using Anti-p47phox Polyclonal Antibody (Product # PA1-9073) and a 50 kDa band corresponding to p47phox was observed across cell lines and tissue extract tested except A-431, PC-3, Mouse Adipose and Mouse Brain which are reported to be low. Membrane enriched extracts (30 µg lysate) of Ramos (Lane 1), Raji (Lane 2), Daudi (Lane 3), A-431 (Lane 4) and PC-3 (Lane 5) and tissue extracts of Mouse adipose (Lane 6), Mouse Brain (Lane 7) and Mouse Spleen (Lane 8) were electrophoresed using Novex® NuPAGE® 4-12 % Bis-Tris gel (Product # NP0322BOX). Resolved proteins were then transferred onto a nitrocellulose membrane (Product # IB23001) by iBlot® 2 Dry Blotting System (Product # IB21001). The blot was probed with the primary antibody (0.5 µg/mL) and detected by chemiluminescence with Rabbit anti-Goat IgG Heavy Chain, Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27014, 1:4000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005).

p47phox Antibody in Western Blot (WB) — Western blot analysis of p47phox using p47phox Polyclonal Antibody (Product # PA1-9073) (0.2 µg/mL) in staining of U937 (A), (0.01 µg/mL) Daudi (B), (0.3 µg/mL) U251 (C) and negative control A431 (D) cell lysate (35 µg protein in RIPA buffer). Detected by chemiluminescence.

p47phox Antibody in Western Blot (WB) — Western blot analysis of p47phox using p47phox Polyclonal Antibody (Product # PA1-9073) (1 µg/mL) in staining of Mouse Thymus (A) and Pig Spleen (B) lysate (35 µg protein in RIPA buffer). Detected by chemiluminescence.

p47phox Antibody in Flow Cytometry (Flow) — Flow cytometric analysis of p47phox in HeLa cells using a polyclonal antibody (Product #PA1-9073). HeLa cells (blue line) were paraformaldehyde fixed and permeabilized with 0.5% Triton. The primary antibody was incubated for one hour (10 µg/mL) followed by an Alexa Fluor 488 secondary antibody (1 µg/mL). IgG control: Unimmunized goat IgG (black line) followed by an Alexa Fluor 488 secondary antibody.

p47phox Antibody in Western Blot (WB) — Figure 5. BOOST of the NCF1 Kozak sequence. ( A ) Schematic representation of the NCF1 wild-type (WT), variant 2 (Var 2) and variant 4 (Var 4) Kozak sequences. The starting codon is bold blue; the base changes in the variants are highlighted in pink. ( B ) Editing efficiency in the Raji bulk population at target and bystander (in red) guanines analysed with the EditR software five days post-electroporation of AncBE4max and sgNCF1 or sgCTRL. The percentage of corrected G-to-A conversions (y-axis) is shown for each position in the NCF1 Kozak sequence (x-axis, with the A of ATG being position + 1). Data are means +- SD from n = 3 independent experiments. ( C ). Editing efficiency in the two clones isolated from the bulk population (Var 2 and Var 4 cells) at target and bystander (in red) guanines. ( D ) Sanger sequencing chromatograms of NCF1 Kozak sequence in Raji WT, Var 2 and Var 4 cells. ( E ) Western blot analysis of the p47 phox protein in Raji cells (WT, Var 2, and Var 4). One representative blot result is shown. The arrow indicates the 47KDa band corresponding to p47 phox . ( F ) Western blot quantification. p47 phox levels were normalised on the housekeeping protein, and the fold change with respect to the WT levels is shown, n = 3 biological replicates. ( G ) qPCR of NCF1 on WT, Var 2 or Var 4 Raji cells. Data are means +- SD from n = 3 independent experiments. ( H ). Representative western blot of two polysomal markers (RPS6 and RPL26) in the fractions isolated by sucr

p47phox Antibody in Western Blot (WB) — 6 FIGURE TRPM2 interacted with p47 phox to induce autophagy and HBV replication in vivo. Mice were simultaneously injected via caudal veins with pHBV1.3 and TRPM2 shRNA (sh-TRPM2), with pNC or NC shRNA (sh-NC) as the control. (A, B): mRNA levels of TRPM2 and p47 phox were determined via RT-qPCR; (C): Protein levels of TRPM2, p47 phox, LC3, Beclin-1 and p62 were measured via Western blot analysis; (D): HBV DNA load in serum was determined via quantitative PCR; (E): Secretion of HBeAg and HBsAg in serum was quantified via ELISA. Experiments were performed 3 times independently. Data in Figures C-E were represented as mean +- SD. Multi-group comparisons in panels C-E were analysed using two-way ANOVA, in panels A, B and D were analysed using one-way ANOVA, followed by Tukey's multiple comparison test. ** p < .01

p47phox Antibody in Western Blot (WB) — 6 Figure Protein expressions of nitrotyrosine and p47phox subunit of NADPH oxidase were increased in the SCD compared to WT mouse penis and were decreased in the SCD mouse penis by FGIN-1-27 treatment. Representative western blots and quantification of nitrotyrosine (a) and p47phox (b) in penes isolated from WT and SCD mice treated with a TSPO ligand FGIN-1-27 or vehicle for nine days. Protein levels of nitrotyrosine and p47phox were analyzed by immunoblotting and normalized to GAPDH expression. Statistical analysis was performed by using one-way analysis of variance, followed by Newman-Keuls multiple comparison tests (between treatment groups) and by modified t test (between control and each treatment group). Data represent mean +- SEM ( n = 5-6). * p < .05 versus WT; # p < .05 versus SCD. FGIN-1-27, N , N -dihexyl-2-(4-fluorophenyl)indole-3-acetamide; GAPDH, glyceraldehyde-3-phosphate dehydrogenase; SCD, sickle cell disease; TSPO, translocator protein; WT, wild-type

p47phox Antibody — Antibody specificity was demonstrated by detection of differential basal expression of the target across cell lysates and tissue extracts tested owing to their inherent genetic constitution. Relative expression of p47phox was observed in Ramos, Raji, Daudi and Mouse Spleen as compared to A-431, PC-3, Mouse Adipose and Mouse Brain using p47phox Polyclonal Antibody (Product # PA1-9073) in Western Blot. {RE}

Product Details

PA1-9073

Applications

Tested Dilution

Publications

Western Blot (WB)

0.01-1 µg/mL

View 3 publications 3 publications

Immunohistochemistry (Paraffin) (IHC (P))

5 µg/mL

-

Immunocytochemistry (ICC/IF)

10 µg/mL

-

Flow Cytometry (Flow)

10 µg/mL

-

Product Specifications

Species Reactivity

Human, Mouse, Pig

Published species

Human, Mouse

Host/Isotype

Goat / IgG

Class

Polyclonal

Type

Antibody

Immunogen

Synthetic peptide corresponding to the C terminus amino acids SESTKRKLASAV (aa 379-390).

Conjugate

Unconjugated

Form

Liquid

Concentration

0.5 mg/mL

Purification

Ammonium sulfate precipitation

Storage buffer

TBS, pH 7.3, with 0.5% BSA

Contains

0.02% sodium azide

Storage conditions

-20°C, Avoid Freeze/Thaw Cycles

Shipping conditions

Ambient (domestic); Wet ice (international)

AB_2150280

Product Specific Information

This antibody is predicted to react with bovine, mouse, porcine, rabbit and rat based on sequence homology.

This antibody is tested in Peptide ELISA: antibody detection limit dilution 20,000.

Target Information

p47phox (NCF1) is required, along with NCF2 and membrane bound cytochrome b558, for activation of the latent NADPH oxidase. This oxidase is a multicomponent enzyme that is activated to produce superoxide anion. Mutations in the gene can result in granulomatous disease.

For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.

Product References

TRPM2 regulates autophagy to participate in hepatitis B virus replication.

Journal of viral hepatitis

Chen L,Zhu L,Lu X,Ming X,Yang B

Published figure using p47phox polyclonal antibody (Product # PA1-9073) in Western Blot

Mon Aug 01 00:00:00 UTC 2022

TRPM2 regulates autophagy to participate in hepatitis B virus replication.

Journal of viral hepatitis

Chen L,Zhu L,Lu X,Ming X,Yang B

Published figure using p47phox polyclonal antibody (Product # PA1-9073) in Western Blot

Mon Aug 01 00:00:00 UTC 2022

Translational enhancement by base editing of the Kozak sequence rescues haploinsufficiency.

Nucleic acids research

Ambrosini C,Destefanis E,Kheir E,Broso F,Alessandrini F,Longhi S,Battisti N,Pesce I,Dassi E,Petris G,Cereseto A,Quattrone A

Published figure using p47phox polyclonal antibody (Product # PA1-9073) in Western Blot

Fri Oct 14 00:00:00 UTC 2022

TSPO ligand FGIN-1-27 controls priapism in sickle cell mice via endogenous testosterone production.

Journal of cellular physiology

Musicki B,Karakus S,La Favor JD,Chen H,Silva FH,Sturny M,Zirkin BR,Burnett AL

Published figure using p47phox polyclonal antibody (Product # PA1-9073) in Western Blot

Thu Apr 01 00:00:00 UTC 2021

TSPO ligand FGIN-1-27 controls priapism in sickle cell mice via endogenous testosterone production.

Journal of cellular physiology

Musicki B,Karakus S,La Favor JD,Chen H,Silva FH,Sturny M,Zirkin BR,Burnett AL

Published figure using p47phox polyclonal antibody (Product # PA1-9073) in Western Blot

Thu Apr 01 00:00:00 UTC 2021

Bioinformatics

Protein Aliases: 47 kDa autosomal chronic granulomatous disease protein; 47 kDa neutrophil oxidase factor; NADPH oxidase organizer 2; NADPH oxidase subunit (47 kD); NADPH oxidase subunit (47kDa); NCF-1; NCF-47K; neutrop; Neutrophil cytosol factor 1; neutrophil cytosolic factor 1, (chronic granulomatous disease, autosomal 1); Neutrophil NADPH oxidase factor 1; Nox organizer 2; Nox-organizing protein 2; P47 phox; p47-phox; SH3 and PX domain-containing protein 1A

Gene Aliases: Ncf-1; NCF-47K; NCF1; NCF1A; NOXO2; p47-phox; p47; p47phox; SH3PXD1A

UniProt ID: (Human) P14598, (Mouse) Q09014

Entrez Gene ID: (Pig) 100134857, (Human) 653361, (Mouse) 17969

Function(s)

protein binding electron carrier activity superoxide-generating NADPH oxidase activity superoxide-generating NADPH oxidase activator activity SH3 domain binding phosphatidylinositol binding phosphatidylinositol-3,4-bisphosphate binding lipid binding

Process(es)

response to yeast leukocyte mediated cytotoxicity respiratory burst involved in defense response protein targeting to membrane leukotriene metabolic process NADP catabolic process apoptotic process inflammatory response cellular defense response cell proliferation response to bacterium superoxide anion generation defense response to bacterium respiratory burst negative regulation of smooth muscle contraction hydrogen peroxide biosynthetic process defense response to Gram-positive bacterium defense response to fungus oxidation-reduction process neutrophil mediated killing of gram-positive bacterium neutrophil mediated killing of fungus antigen processing and presentation of exogenous peptide antigen via MHC class I, TAP-dependent superoxide metabolic process positive regulation of catalytic activity innate immune response cell redox homeostasis vascular endothelial growth factor receptor signaling pathway

Performance Guarantee

If an Invitrogen™ antibody doesn't perform as described on our website or datasheet,we'll replace the product at no cost to you, or provide you with a credit for a future purchase.*

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