Cytochrome B245 antibody | NL7

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Mouse anti Human Cytochrome B245 Heavy Chain:Alexa Fluor® 488

Mouse anti Human Cytochrome B245 Heavy Chain:Alexa Fluor® 647

Mouse anti Human Cytochrome B245 Heavy Chain

Product Type
Monoclonal Antibody
Clone
NL7
Isotype
IgG1
Product CodeApplicationsDatasheetMSDSPack SizeList PriceQuantity
MCA4685A488 F* 100 Tests/1ml
MCA4685A647 F* 100 Tests/1ml
MCA4685GA C E F* IF WB 0.1 mg
MCA4685 C E F* IF WB 0.2 mg
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.

Product Details

Target Species
Human
Product Form
Purified IgG conjugated to Alexa Fluor 488 - liquid
Product Form
Purified IgG conjugated to Alexa Fluor® 647 - liquid
Product Form
Purified IgG - liquid
Preparation
Purified IgG prepared by affinity chromatography on Protein G from tissue culture supernatant
Preparation
Purified IgG prepared by affinity chromatography on Protein G from tissue culture supernatant
Preparation
Purified IgG prepared by affinity chromatography on Protein G from tissue culture supernatant
Buffer Solution
Phosphate buffered saline
Buffer Solution
Phosphate buffered saline
Buffer Solution
Phosphate buffered saline
Preservative Stabilisers
0.09% Sodium Azide (NaN3)
1% Bovine Serum Albumin
Preservative Stabilisers
0.09% Sodium Azide (NaN3)
1% Bovine Serum Albumin
Preservative Stabilisers
0.09% Sodium Azide (NaN3)
Carrier Free
Yes
Immunogen
Cytochrome B245 solubilised in octyl-beta-glucopyranoside.
Approx. Protein Concentrations
IgG concentration 0.05mg/ml
Approx. Protein Concentrations
IgG concentration 0.05mg/ml
Approx. Protein Concentrations
IgG concentration 1.0mg/ml
Fusion Partners
Spleen cells from immunised BALB/c were fused with cells of the 3PU1 myeloma cell line.

Storage Information

Storage
Store at +4oC or at -20oC if preferred.
Storage in frost-free freezers is not recommended.
This product should be stored undiluted. This product is photosensitive and should be protected from light. Avoid repeated freezing and thawing as this may denature the antibody. Should this product contain a precipitate we recommend microcentrifugation before use.
Storage
Store at +4oC or at -20oC if preferred.
Storage in frost-free freezers is not recommended.
This product should be stored undiluted. This product is photosensitive and should be protected from light. Avoid repeated freezing and thawing as this may denature the antibody. Should this product contain a precipitate we recommend microcentrifugation before use.
Storage
Store at +4oC or at -20oC if preferred.
Storage in frost-free freezers is not recommended.
This product should be stored undiluted. Avoid repeated freezing and thawing as this may denature the antibody. Should this product contain a precipitate we recommend microcentrifugation before use.
Shelf Life
18 months from date of despatch.
Shelf Life
18 months from date of despatch.
Shelf Life
18 months from date of despatch.

More Information

UniProt
P04839 Related reagents
Entrez Gene
CYBB Related reagents
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
Acknowledgements
This product is provided under an intellectual property licence from Life Technologies Corporation. The transfer of this product is contingent on the buyer using the purchase product solely in research, excluding contract research or any fee for service research, and the buyer must not sell or otherwise transfer this product or its components for (a) diagnostic, therapeutic or prophylactic purposes; (b) testing, analysis or screening services, or information in return for compensation on a per-test basis; (c) manufacturing or quality assurance or quality control, or (d) resale, whether or not resold for use in research. For information on purchasing a license to this product for purposes other than as described above, contact Life Technologies Corporation, 5791 Van Allen Way, Carlsbad CA 92008 USA or outlicensing@thermofisher.com
Acknowledgements
This product is provided under an intellectual property licence from Life Technologies Corporation. The transfer of this product is contingent on the buyer using the purchase product solely in research, excluding contract research or any fee for service research, and the buyer must not sell or otherwise transfer this product or its components for (a) diagnostic, therapeutic or prophylactic purposes; (b) testing, analysis or screening services, or information in return for compensation on a per-test basis; (c) manufacturing or quality assurance or quality control, or (d) resale, whether or not resold for use in research. For information on purchasing a license to this product for purposes other than as described above, contact Life Technologies Corporation, 5791 Van Allen Way, Carlsbad CA 92008 USA or outlicensing@thermofisher.com
Regulatory
For research purposes only

Applications of Cytochrome B245 antibody

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
Flow Cytometry 1 Neat
Flow Cytometry 1 Neat
ELISA
Flow Cytometry 1 1/25 1/200
Immunofluorescence
Immunohistology - Frozen
Western Blotting
  1. 1Membrane permeabilisation is required for this application. Bio-Rad recommends the use of Leucoperm™ (Product Code BUF09) for this purpose.
  1. 1Membrane permeabilisation is required for this application. Bio-Rad recommends the use of Leucoperm™ (Product Code BUF09) for this purpose.
  1. 1Membrane permeabilisation is required for this application. Bio-Rad recommends the use of Leucoperm™ (Product Code BUF09) 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.
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.
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.
Flow Cytometry
Use 10ul of the suggested working dilution to label 1x106 cells in 100ul.
Flow Cytometry
Use 10ul of the suggested working dilution to label 1x106 cells in 100ul.
Western Blotting
MCA4685 detects a broad band of approximately 90kDa

Secondary Antibodies Available

Description Product Code Pack Size Applications List Price Quantity
Human anti Mouse IgG1:HRP HCA036P 0.1 mg E
Goat anti Mouse IgG (H/L):Alk. Phos. (Multi Species Adsorbed) STAR117A 0.5 mg E WB
Goat anti Mouse IgG (H/L):DyLight®488 (Multi Species Adsorbed) STAR117D488GA 0.1 mg F IF
Goat anti Mouse IgG (H/L):DyLight®549 (Multi Species Adsorbed) STAR117D549GA 0.1 mg F IF WB
Goat anti Mouse IgG (H/L):DyLight®649 (Multi Species Adsorbed) STAR117D649GA 0.1 mg F IF
Goat anti Mouse IgG (H/L):DyLight®680 (Multi Species Adsorbed) STAR117D680GA 0.1 mg F WB
Goat anti Mouse IgG (H/L):DyLight®800 (Multi Species Adsorbed) STAR117D800GA 0.1 mg F IF WB
Goat anti Mouse IgG (H/L):FITC (Multi Species Adsorbed) STAR117F 0.5 mg F
Goat anti Mouse IgG (H/L):HRP (Multi Species Adsorbed) STAR117P 0.5 mg E WB
Goat anti Mouse IgG (Fc):FITC STAR120F 1 mg C F
Goat anti Mouse IgG (Fc):HRP STAR120P 1 mg E WB
Rabbit F(ab')2 anti Mouse IgG:RPE STAR12A 1 ml F
Rabbit F(ab')2 anti Mouse IgG:HRP (Human Adsorbed) STAR13B 1 mg C E P RE WB
Goat anti Mouse IgG:FITC (Rat Adsorbed) STAR70 0.5 mg F
Goat anti Mouse IgG:RPE (Rat Adsorbed) STAR76 1 ml F
Goat anti Mouse IgG:HRP (Rat Adsorbed) STAR77 0.5 mg C E P
Goat anti Mouse IgG/A/M:Alk. Phos. STAR87A 1 mg C E WB
Goat anti Mouse IgG/A/M:HRP (Human Adsorbed) STAR87P 1 mg E
Rabbit F(ab')2 anti Mouse IgG:Dylight®800 STAR8D800GA 0.1 mg F IF WB
Rabbit F(ab')2 anti Mouse IgG:FITC STAR9B 1 mg F

Negative Isotype Controls Available

Description Product Code Pack Size Applications List Price Quantity
Mouse IgG1 Negative Control:Alexa Fluor® 488 MCA928A488 100 Tests/1ml F
Mouse IgG1 Negative Control:Alexa Fluor® 647 MCA928A647 100 Tests/1ml F
Mouse IgG1 Negative Control MCA928 100 Tests F

Useful Reagents Available

Description Product Code Pack Size Applications List Price Quantity
Human Seroblock BUF070A 50 Test F
Human Seroblock BUF070B 200 Test F
Human Seroblock BUF070A 50 Test F
Human Seroblock BUF070B 200 Test F

Application Based External Images

Immunofluorescence

Product Specific References

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. (2010) Fc gamma receptors are required for NF-kappaB 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.