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Cytochrome B245 antibody | NL7

Mouse anti Human Cytochrome B245 Heavy Chain

Product Type
Monoclonal Antibody
Clone
NL7
Isotype
IgG1
Specificity
Cytochrome B245
Region
HEAVY CHAIN

Product Code Applications Pack Size List Price Your Price Qty
MCA4685GA
Datasheet Datasheet Datasheet
SDS Safety Datasheet SDS
C E F * IF WB 0.1 mg loader
List Price Your Price
loader
MCA4685
Datasheet Datasheet Datasheet
SDS Safety Datasheet SDS
C E F * IF WB 0.2 mg loader
List Price Your Price
loader

Mouse anti Human Cytochrome B245 Heavy Chain antibody, clone NL7 recognizes the heavy chain of cytochrome B245, also known as gp91phox or NOX2, a subunit of the nicotinamide adenine dinucleotide phosphate-oxidase (NADPH oxidase). NADPH oxidase is a membrane-bound enzyme complex in neutrophils that produces the superoxide anion (O2-) by transferring single electrons from NADPH inside the cell across the membrane and coupling them to molecular oxygen. Superoxide is used for the destruction of pathogens. Individuals with defects in NADPH oxidase suffer from chronic granulomatous disease, characterised by severe recurrent infections.

The heavy chain (gp91phox), along with the light chain (p22phox) form cytochrome B245 (also known as flavocytochome b, or Cyt b), which is always membrane-resident. Upon the appropriate stimulus, other components of NADPH oxidase (p47phox, p40phox, p67phox and Rac2) rapidly translocate from the cytosol to the cell membrane to form the complete enzyme complex. Cytochrome B245 appears to serve as a scaffold for the assembly of the other cytosolic units of the oxidase and to provide a transmembrane pathway for electrons.

Mouse anti Human Cytochrome B245 Heavy Chain antibody, clone NL7 binds amino acids 498-506 (EKDVITGLK), a cytosolic epitope on gp91phox not accessible on intact neutrophils. It recognizes the native as well as the denatured form of the protein. Clone NL7 has an inhibitory effect on NADPH oxidase activation. Research indicates that this inhibitory effect is exerted during the initial stages of oxidase activation. The antibody does not interfere with the translocation of the cytosolic subunits of NADPH oxidase to the cell membrane.

Target Species
Human
Product Form
Purified IgG - liquid
Preparation
MCA4685GA: Purified IgG prepared by affinity chromatography on Protein A from tissue culture supernatant
MCA4685: Purified IgG prepared by affinity chromatography on Protein G from tissue culture supernatant.
Buffer Solution
MCA4685GA: Phosphate buffered saline
MCA4685: Phosphate buffered saline.
Preservative Stabilisers
MCA4685GA: 0.09% Sodium Azide (NaN3)
MCA4685: 0.09% Sodium Azide (NaN3).
Carrier Free
Yes
Immunogen
Cytochrome B245 solubilized in octyl-beta-glucopyranoside.
Approx. Protein Concentrations
MCA4685GA: IgG concentration 1.0mg/ml
MCA4685: IgG concentration 1.0mg/ml.
Fusion Partners
Spleen cells from immunized BALB/c were fused with cells of the 3PU1 myeloma cell line.
Regulatory
MCA4685GA: For research purposes only
MCA4685: For research purposes only.
Guarantee
MCA4685GA: 12 months from date of despatch
MCA4685: 12 months from date of despatch.

This product is shipped at ambient temperature. It is recommended to aliquot and store at -20°C on receipt. When thawed, aliquot the sample as needed. Keep aliquots at 2-8°C for short term use (up to 4 weeks) and store the remaining aliquots at -20°C.

Avoid repeated freezing and thawing as this may denature the antibody. Storage in frost-free freezers is not recommended.

This product has been reported to work in the following applications. This information is derived from testing within our laboratories, peer-reviewed publications or personal communications from the originators. Please refer to references indicated for further information. For general protocol recommendations, please visit the antibody protocols page.
Application Name Verified Min Dilution Max Dilution
ELISA
Flow Cytometry 1 1/25 1/200
Immunofluorescence
Immunohistology - Frozen
Western Blotting
  1. 1 Membrane permeabilization is required for this application. The use of Leucoperm (Product Code BUF09) is recommended for this purpose.
Where this product has not been tested for use in a particular technique this does not necessarily exclude its use in such procedures. Suggested working dilutions are given as a guide only. It is recommended that the user titrates the product for use in their own system using appropriate negative/positive controls.
Flow Cytometry
Use 10ul of the suggested working dilution to label 1x106 cells in 100ul.
Western Blotting
Mouse anti Human cytochrome B245 heavy chain antibody, clone NL7 detects a broad band of approximately 90kDa.

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Description Goat anti Mouse IgG (H/L):DyLight®650 (Multi Species Adsorbed)
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Description Goat anti Mouse IgG (H/L):DyLight®680 (Multi Species Adsorbed)
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Description Goat anti Mouse IgG (H/L):DyLight®800 (Multi Species Adsorbed)
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Description Goat anti Mouse IgG (H/L):FITC (Multi Species Adsorbed)
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Description Goat anti Mouse IgG (H/L):HRP (Multi Species Adsorbed)
Goat anti Mouse IgG (Fc):FITC STAR120F C F 1 mg loader
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Description Goat anti Mouse IgG (Fc):FITC
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Description Goat anti Mouse IgG (Fc):HRP
Rabbit F(ab')2 anti Mouse IgG:RPE STAR12A F 1 ml loader
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Description Rabbit F(ab')2 anti Mouse IgG:RPE
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Description Rabbit F(ab')2 anti Mouse IgG:HRP (Human Adsorbed)
Goat anti Mouse IgG:FITC (Rat Adsorbed) STAR70 F 0.5 mg loader
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Description Goat anti Mouse IgG:FITC (Rat Adsorbed)
Goat anti Mouse IgG:RPE (Rat Adsorbed) STAR76 F 1 ml loader
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Description Goat anti Mouse IgG:RPE (Rat Adsorbed)
Goat anti Mouse IgG:HRP (Rat Adsorbed) STAR77 C E P 0.5 mg loader
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Description Goat anti Mouse IgG:HRP (Rat Adsorbed)
Goat anti Mouse IgG/A/M:HRP (Human Adsorbed) STAR87P E 1 mg loader
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Description Goat anti Mouse IgG/A/M:HRP (Human Adsorbed)
Rabbit F(ab')2 anti Mouse IgG:FITC STAR9B F 1 mg loader
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Description Rabbit F(ab')2 anti Mouse IgG:FITC

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Mouse IgG1 Negative Control MCA928 F 100 Tests loader
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Description Mouse IgG1 Negative Control

References for Cytochrome B245 antibody

  1. Burritt, J.B. et al. (2003) Functional epitope on human neutrophil flavocytochrome b558.
    J Immunol. 170 (12): 6082-9.
  2. Taylor, R.M. et al. (2004) Site-specific inhibitors of NADPH oxidase activity and structural probes of flavocytochrome b: characterization of six monoclonal antibodies to the p22phox subunit.
    J Immunol. 173 (12): 7349-57.
  3. Taylor, R.M. et al. (2007) Cloning, sequence analysis and confirmation of derived gene sequences for three epitope-mapped monoclonal antibodies against human phagocyte flavocytochrome b.
    Mol Immunol. 44 (4): 625-37.
  4. Cao, S. et al. (2010) Fcγ receptors are required for NF-κB signaling, microglial activation and dopaminergic neurodegeneration in an AAV-synuclein mouse model of Parkinson's disease.
    Mol Neurodegener. 5: 42.
  5. Kovács I et al. (2015) Reactive oxygen species-mediated bacterial killing by B lymphocytes.
    J Leukoc Biol. 97 (6): 1133-7.

Immunofluorescence

Synonyms
gp91phox
RRID
AB_1963543
UniProt
P04839
Entrez Gene
CYBB
GO Terms
GO:0009055 electron carrier activity
GO:0005244 voltage-gated ion channel activity
GO:0022900 electron transport chain
GO:0006954 inflammatory response
GO:0016175 superoxide-generating NADPH oxidase activity
GO:0020037 heme binding
GO:0042554 superoxide anion generation
GO:0043020 NADPH oxidase complex
GO:0045087 innate immune response
GO:0045730 respiratory burst
GO:0046982 protein heterodimerization activity
GO:0050660 flavin adenine dinucleotide binding

MCA4685GA

150626 167503

MCA4685

154534 1612 162884

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