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p47-phox Mouse Monoclonal Antibody [A6B11]

Usd: 350

Specification

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Catalog# HA600049

p47-phox Mouse Monoclonal Antibody [A6B11]

Application

  • WB

  • IF-Cell

  • IHC-P

  • FC

Reactivity

  • Human

  • Mouse

Conjugation

  • unconjugated

This product has been cited in peer reviewed publications, see list HERE

Overview

Product Name

p47-phox Mouse Monoclonal Antibody [A6B11]

Antibody Type

Mouse Monoclonal Antibody

Immunogen

Recombinant protein within human p47-phox aa 51-250.

Species Reactivity

Human, Mouse

Validated Applications

WB, IF-Cell, IHC-P, FC

Molecular Weight

Predicted band size: 45 kDa

Positive Control

Daudi cell lysates, Hela, NIH/3T3, human lung tissue, human spleen tissue, Daudi.

Conjugation

unconjugated

Clone Number

A6B11

RRID

AB_3071666

Product Features

Form

Liquid

Concentration

Storage Instructions

Shipped at 4℃. Store at +4℃ short term (1-2 weeks). Store at -20℃ long term.

Storage Buffer

PBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.

Isotype

IgG1

Purification Method

Protein G affinity purified.

Application Dilution

  • WB

  • 1:2,000-1:5,000

  • IF-Cell

  • 1:50-1:200

  • IHC-P

  • 1:1,000

  • FC

  • 1:100

Target

Function

The heredity chronic granulomatous disease (CGF) has been linked to mutations in p47phox and p67-phox. The cytosolic proteins p47phox and p67-phox, also designated neutrophil cytosol factor (NCF)1 and NCF2, respectively, are required for activation of the superoxide-producing NADPH oxidase in neutrophils and other phagocytic cells. During activation of the NADPH oxidase, p47phox and p67-phox migrate to the plasma membrane where they associate with cytochrome b558 and the small G protein Rac to form the functional enzyme complex. Both p47phox and p67-phox contain two Src homology 3 (SH3) domains. The C-terminal SH3 domain of p67-phox has been shown to interact with the proline-rich domain of p47phox, suggesting that p47phox may faciliate the transport of p67-phox to the membrane.

Background References

1. Gong P. et. al. p47(phox) deficiency improves cognitive impairment and attenuates tau hyperphosphorylation in mouse models of AD. Alzheimers Res Ther. 2020 Nov

2. Kuhns DB. et. al. NCF1 (p47(phox))-deficient chronic granulomatous disease: comprehensive genetic and flow cytometric analysis. Blood Adv. 2019 Jan

Subcellular Location

Cytoplasm, cytosol, Membrane.

UNIPROT

SwissProt: P14598 Human

SwissProt: Q09014 Mouse

Synonyms

47 kDa autosomal chronic granulomatous disease protein antibody

47 kDa neutrophil oxidase factor antibody

NADPH oxidase organizer 2 antibody

NCF 47K antibody

NCF-1 antibody

NCF-47K antibody

Ncf1 antibody

NCF1_HUMAN antibody

Neutrophil cytosol factor 1 antibody

Neutrophil cytosolic factor 1 antibody

Expand

Images

  • Western blot analysis of p47-phox on Daudi cell lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (<a href="/products/HA600049" style="font-weight: bold;text-decoration: underline;">HA600049</a>, 1/500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Mouse IgG - HRP Secondary Antibody (<a href="/products/HA1006" style="font-weight: bold;text-decoration: underline;">HA1006</a>) at 1:20,000 dilution was used for 1 hour at room temperature.

    Western blot analysis of p47-phox on Daudi cell lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (HA600049, 1/500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Mouse IgG - HRP Secondary Antibody (HA1006) at 1:20,000 dilution was used for 1 hour at room temperature.

  • ICC staining of p47-phox in Hela cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 10% negative goat serum for 15 minutes at room temperature. Cells were probed with the primary antibody (<a href="/products/HA600049" style="font-weight: bold;text-decoration: underline;">HA600049</a>, 1/100) for 1 hour at room temperature, washed with PBS. Alexa Fluor&reg;488 conjugate-Goat anti-Mouse IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).

    ICC staining of p47-phox in Hela cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 10% negative goat serum for 15 minutes at room temperature. Cells were probed with the primary antibody (HA600049, 1/100) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 conjugate-Goat anti-Mouse IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).

  • ICC staining of p47-phox in NIH/3T3 cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 10% negative goat serum for 15 minutes at room temperature. Cells were probed with the primary antibody (<a href="/products/HA600049" style="font-weight: bold;text-decoration: underline;">HA600049</a>, 1/100) for 1 hour at room temperature, washed with PBS. Alexa Fluor&reg;488 conjugate-Goat anti-Mouse IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).

    ICC staining of p47-phox in NIH/3T3 cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 10% negative goat serum for 15 minutes at room temperature. Cells were probed with the primary antibody (HA600049, 1/100) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 conjugate-Goat anti-Mouse IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).

  • Immunohistochemical analysis of paraffin-embedded human lung tissue using anti-p47-phox antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes.The tissues were blocked in 1% BSA for 30 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/HA600049" style="font-weight: bold;text-decoration: underline;">HA600049</a>, 1/1,000) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

    Immunohistochemical analysis of paraffin-embedded human lung tissue using anti-p47-phox antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes.The tissues were blocked in 1% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA600049, 1/1,000) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

  • Immunohistochemical analysis of paraffin-embedded human spleen tissue using anti-p47-phox antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes.The tissues were blocked in 1% BSA for 30 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/HA600049" style="font-weight: bold;text-decoration: underline;">HA600049</a>, 1/1,000) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

    Immunohistochemical analysis of paraffin-embedded human spleen tissue using anti-p47-phox antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes.The tissues were blocked in 1% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA600049, 1/1,000) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

  • Flow cytometric analysis of p47-phox was done on Daudi cells. The cells were fixed, permeabilized and stained with the primary antibody (<a href="/products/HA600049" style="font-weight: bold;text-decoration: underline;">HA600049</a>, 10ug/ml) (red) compared with Rabbit IgG, monoclonal - Isotype Control (green). After incubation of the primary antibody at +4℃ for 1 hour, the cells were stained with a Alexa Fluor&reg;488 conjugate-Goat anti-Mouse IgG Secondary antibody at 1/1000 dilution for 30 minutes at +4℃ (dark incubation).Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

    Flow cytometric analysis of p47-phox was done on Daudi cells. The cells were fixed, permeabilized and stained with the primary antibody (HA600049, 10ug/ml) (red) compared with Rabbit IgG, monoclonal - Isotype Control (green). After incubation of the primary antibody at +4℃ for 1 hour, the cells were stained with a Alexa Fluor®488 conjugate-Goat anti-Mouse IgG Secondary antibody at 1/1000 dilution for 30 minutes at +4℃ (dark incubation).Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

Citation