Western Blot (WB)
(Figure 1. Western blot analysis of NOX2/gp91phox/CYBB using anti-NOX2/gp91phox/CYBB antibody (MBS1753313).Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 50ug of sample under reducing conditions.Lane 1: human HELA whole cell lysatesLane 2: human HEPG2 whole cell lysatesLane 3: human U937 whole cell lysates.After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA for 50-90 minutes. Blocked the membrane with 5% Non-fat Milk/ TBS for 1. 5 hour at RT. The membrane was incubated with rabbit anti-NOX2/gp91phox/CYBB antigen affinity purified polyclonal antibody (Catalog # MBS1753313) at 0. 5 μg/mL overnight at 4 degree C, then washed with TBS-0. 1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:5000 for 1. 5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # MBS176460) with Tanon 5200 system. A specific band was detected for NOX2/gp91phox/CYBB at approximately 65KD. The expected band size for NOX2/gp91phox/CYBB is at 65KD. )
Rabbit anti-Human NOX2/gp91phox/CYBB Polyclonal Antibody | anti-CYBB antibody
Anti-NOX2/gp91phox/CYBB Antibody
IHC-P: 2-5ug/ml|Human|
ICC/IF: 5ug/ml|Human|
FC/FACS/FCM: 1-3ug/1x106 cells|Human|
Direct ELISA: 0.1-0.5ug/ml|Human|
Western Blot (WB)
(Figure 1. Western blot analysis of NOX2/gp91phox/CYBB using anti-NOX2/gp91phox/CYBB antibody (MBS1753313).Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 50ug of sample under reducing conditions.Lane 1: human HELA whole cell lysatesLane 2: human HEPG2 whole cell lysatesLane 3: human U937 whole cell lysates.After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA for 50-90 minutes. Blocked the membrane with 5% Non-fat Milk/ TBS for 1. 5 hour at RT. The membrane was incubated with rabbit anti-NOX2/gp91phox/CYBB antigen affinity purified polyclonal antibody (Catalog # MBS1753313) at 0. 5 μg/mL overnight at 4 degree C, then washed with TBS-0. 1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:5000 for 1. 5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # MBS176460) with Tanon 5200 system. A specific band was detected for NOX2/gp91phox/CYBB at approximately 65KD. The expected band size for NOX2/gp91phox/CYBB is at 65KD. )
Western Blot (WB)
(Figure 1. Western blot analysis of NOX2/gp91phox/CYBB using anti-NOX2/gp91phox/CYBB antibody (MBS1753313).Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 50ug of sample under reducing conditions.Lane 1: human HELA whole cell lysatesLane 2: human HEPG2 whole cell lysatesLane 3: human U937 whole cell lysates.After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA for 50-90 minutes. Blocked the membrane with 5% Non-fat Milk/ TBS for 1. 5 hour at RT. The membrane was incubated with rabbit anti-NOX2/gp91phox/CYBB antigen affinity purified polyclonal antibody (Catalog # MBS1753313) at 0. 5 μg/mL overnight at 4 degree C, then washed with TBS-0. 1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:5000 for 1. 5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # MBS176460) with Tanon 5200 system. A specific band was detected for NOX2/gp91phox/CYBB at approximately 65KD. The expected band size for NOX2/gp91phox/CYBB is at 65KD. )
Immunohistochemistry (IHC)
(Figure 2. IHC analysis of NOX2/gp91phox/CYBB using anti-NOX2/gp91phox/CYBB antibody (MBS1753313).NOX2/gp91phox/CYBB was detected in paraffin-embedded section of human appendicitis tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8. 0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2μg/ml rabbit anti-NOX2/gp91phox/CYBB Antibody (MBS1753313) overnight at 4 degree C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # MBS176451) with DAB as the chromogen. )
Immunohistochemistry (IHC)
(Figure 2. IHC analysis of NOX2/gp91phox/CYBB using anti-NOX2/gp91phox/CYBB antibody (MBS1753313).NOX2/gp91phox/CYBB was detected in paraffin-embedded section of human appendicitis tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8. 0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2μg/ml rabbit anti-NOX2/gp91phox/CYBB Antibody (MBS1753313) overnight at 4 degree C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # MBS176451) with DAB as the chromogen. )
Immunohistochemistry (IHC)
(Figure 3. IHC analysis of NOX2/gp91phox/CYBB using anti-NOX2/gp91phox/CYBB antibody (MBS1753313).NOX2/gp91phox/CYBB was detected in paraffin-embedded section of human bladder cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8. 0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2μg/ml rabbit anti-NOX2/gp91phox/CYBB Antibody (MBS1753313) overnight at 4 degree C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # MBS176451) with DAB as the chromogen. )
Immunohistochemistry (IHC)
(Figure 3. IHC analysis of NOX2/gp91phox/CYBB using anti-NOX2/gp91phox/CYBB antibody (MBS1753313).NOX2/gp91phox/CYBB was detected in paraffin-embedded section of human bladder cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8. 0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2μg/ml rabbit anti-NOX2/gp91phox/CYBB Antibody (MBS1753313) overnight at 4 degree C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # MBS176451) with DAB as the chromogen. )
Immunohistochemistry (IHC)
(Figure 4. IHC analysis of NOX2/gp91phox/CYBB using anti-NOX2/gp91phox/CYBB antibody (MBS1753313).NOX2/gp91phox/CYBB was detected in paraffin-embedded section of human gastric cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8. 0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2μg/ml rabbit anti-NOX2/gp91phox/CYBB Antibody (MBS1753313) overnight at 4 degree C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # MBS176451) with DAB as the chromogen. )
Immunohistochemistry (IHC)
(Figure 4. IHC analysis of NOX2/gp91phox/CYBB using anti-NOX2/gp91phox/CYBB antibody (MBS1753313).NOX2/gp91phox/CYBB was detected in paraffin-embedded section of human gastric cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8. 0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2μg/ml rabbit anti-NOX2/gp91phox/CYBB Antibody (MBS1753313) overnight at 4 degree C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # MBS176451) with DAB as the chromogen. )
Immunohistochemistry (IHC)
(Figure 5. IHC analysis of NOX2/gp91phox/CYBB using anti-NOX2/gp91phox/CYBB antibody (MBS1753313).NOX2/gp91phox/CYBB was detected in paraffin-embedded section of human testicular cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8. 0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2μg/ml rabbit anti-NOX2/gp91phox/CYBB Antibody (MBS1753313) overnight at 4 degree C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # MBS176451) with DAB as the chromogen. )
Immunohistochemistry (IHC)
(Figure 5. IHC analysis of NOX2/gp91phox/CYBB using anti-NOX2/gp91phox/CYBB antibody (MBS1753313).NOX2/gp91phox/CYBB was detected in paraffin-embedded section of human testicular cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8. 0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2μg/ml rabbit anti-NOX2/gp91phox/CYBB Antibody (MBS1753313) overnight at 4 degree C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # MBS176451) with DAB as the chromogen. )
Immunohistochemistry (IHC)
(Figure 6. IHC analysis of NOX2/gp91phox/CYBB using anti-NOX2/gp91phox/CYBB antibody (MBS1753313).NOX2/gp91phox/CYBB was detected in paraffin-embedded section of human melanoma tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8. 0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2μg/ml rabbit anti-NOX2/gp91phox/CYBB Antibody (MBS1753313) overnight at 4 degree C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # MBS176451) with DAB as the chromogen. )
Immunohistochemistry (IHC)
(Figure 6. IHC analysis of NOX2/gp91phox/CYBB using anti-NOX2/gp91phox/CYBB antibody (MBS1753313).NOX2/gp91phox/CYBB was detected in paraffin-embedded section of human melanoma tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8. 0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2μg/ml rabbit anti-NOX2/gp91phox/CYBB Antibody (MBS1753313) overnight at 4 degree C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # MBS176451) with DAB as the chromogen. )
Immunofluorescence (IF)
(Figure 7. IF analysis of NOX2/gp91phox/CYBB using anti- NOX2/gp91phox/CYBB antibody (MBS1753313).NOX2/gp91phox/CYBB was detected in immunocytochemical section of A549 cells. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent (MBS176582) for 15 mins. The cells were blocked with 10% goat serum. And then incubated with 5μg/mL rabbit anti-NOX2/gp91phox/CYBB Antibody (MBS1753313) overnight at 4 degree C. DyLight®488 Conjugated Goat Anti-Rabbit IgG was used as secondary antibody at 1:100 dilution and incubated for 30 minutes at 37 degree C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used. )
Immunofluorescence (IF)
(Figure 7. IF analysis of NOX2/gp91phox/CYBB using anti- NOX2/gp91phox/CYBB antibody (MBS1753313).NOX2/gp91phox/CYBB was detected in immunocytochemical section of A549 cells. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent (MBS176582) for 15 mins. The cells were blocked with 10% goat serum. And then incubated with 5μg/mL rabbit anti-NOX2/gp91phox/CYBB Antibody (MBS1753313) overnight at 4 degree C. DyLight®488 Conjugated Goat Anti-Rabbit IgG was used as secondary antibody at 1:100 dilution and incubated for 30 minutes at 37 degree C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used. )
Flow Cytometry (FC/FACS)
(Figure 8. Flow Cytometry analysis of THP-1 cells using anti-NOX2/gp91phox/CYBB antibody (MBS1753313).Overlay histogram showing THP-1 cells stained with MBS1753313 (Blue line). The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-NOX2/gp91phox/CYBB Antibody (MBS1753313, 1μg/1x106 cells) for 30 min at 20 degree C. DyLight®488 conjugated goat anti-rabbit IgG (5-10μg/1x106 cells) was used as secondary antibody for 30 minutes at 20 degree C. Isotype control antibody (Green line) was rabbit IgG (1μg/1x106) used under the same conditions. Unlabelled sample (Red line) was also used as a control. )
Flow Cytometry (FC/FACS)
(Figure 8. Flow Cytometry analysis of THP-1 cells using anti-NOX2/gp91phox/CYBB antibody (MBS1753313).Overlay histogram showing THP-1 cells stained with MBS1753313 (Blue line). The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-NOX2/gp91phox/CYBB Antibody (MBS1753313, 1μg/1x106 cells) for 30 min at 20 degree C. DyLight®488 conjugated goat anti-rabbit IgG (5-10μg/1x106 cells) was used as secondary antibody for 30 minutes at 20 degree C. Isotype control antibody (Green line) was rabbit IgG (1μg/1x106) used under the same conditions. Unlabelled sample (Red line) was also used as a control. )
2. Bolscher, B. G. J. M., de Boer, M., de Klein, A., Weening, R. S., Roos, D. Point mutations in the beta-subunit of cytochrome b(558) leading to X-linked chronic granulomatous disease. Blood 77: 2482-2487, 1991.
3. "Entrez Gene: CYBB cytochrome b-245, beta polypeptide (chronic granulomatous disease)".
NCBI and Uniprot Product Information
NCBI Description
Cytochrome b (-245) is composed of cytochrome b alpha (CYBA) and beta (CYBB) chain. It has been proposed as a primary component of the microbicidal oxidase system of phagocytes. CYBB deficiency is one of five described biochemical defects associated with chronic granulomatous disease (CGD). In this disorder, there is decreased activity of phagocyte NADPH oxidase; neutrophils are able to phagocytize bacteria but cannot kill them in the phagocytic vacuoles. The cause of the killing defect is an inability to increase the cell's respiration and consequent failure to deliver activated oxygen into the phagocytic vacuole. [provided by RefSeq, Jul 2008]
Uniprot Description
CYBB: Critical component of the membrane-bound oxidase of phagocytes that generates superoxide. It is the terminal component of a respiratory chain that transfers single electrons from cytoplasmic NADPH across the plasma membrane to molecular oxygen on the exterior. Also functions as a voltage-gated proton channel that mediates the H(+) currents of resting phagocytes. It participates in the regulation of cellular pH and is blocked by zinc. Defects in CYBB are a cause of granulomatous disease,chronic, X-linked (CGD). A disorder characterized by the inability of neutrophils and phagocytes to kill microbes that they have ingested. Patients suffer from life- threatening bacterial/fungal infections. Defects in CYBB are a cause of mycobacteriosis atypical X-linked type 2 (AMCBX2). A rare condition characterized by predisposition to illness caused by moderately virulent mycobacterial species, such as Bacillus Calmette-Guerin (BCG) vaccine and environmental non-tuberculous mycobacteria, and by the more virulent Mycobacterium tuberculosis. Other microorganisms rarely cause severe clinical disease in individuals with susceptibility to mycobacterial infections.
Protein type: Oxidoreductase; Mitochondrial; EC 1.-.-.-; Membrane protein, integral; Membrane protein, multi-pass
Chromosomal Location of Human Ortholog: Xp21.1
Cellular Component: Golgi apparatus; phagocytic vesicle membrane; rough endoplasmic reticulum; cell soma; mitochondrion; integral to plasma membrane; dendrite; plasma membrane; NADPH oxidase complex
Molecular Function: protein binding; FAD binding; electron carrier activity; protein heterodimerization activity; metal ion binding; superoxide-generating NADPH oxidase activity; heme binding; voltage-gated ion channel activity
Biological Process: response to drug; respiratory burst; interaction with host; superoxide metabolic process; antigen processing and presentation of exogenous peptide antigen via MHC class I, TAP-dependent; antigen processing and presentation of peptide antigen via MHC class I; innate immune response; antigen processing and presentation of exogenous peptide antigen via MHC class I; ion transport; vascular endothelial growth factor receptor signaling pathway; inflammatory response; superoxide release; hydrogen peroxide biosynthetic process; response to nutrient
Disease: Immunodeficiency 34; Granulomatous Disease, Chronic, X-linked
Research Articles on CYBB
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Product Notes
The CYBB cybb (Catalog #AAA1753313) is an Antibody produced from Rabbit and is intended for research purposes only. The product is available for immediate purchase. The Anti-NOX2/gp91phox/CYBB Antibody reacts with Human and may cross-react with other species as described in the data sheet. AAA Biotech's NOX2/gp91phox/CYBB can be used in a range of immunoassay formats including, but not limited to, Western Blot (WB), Immunohistochemistry-Paraffin (IHC-P), Immunocytochemistry (ICC), Immunofluorescence (IF), Flow Cytometry (FC/FACS/FCM), Direct ELISA (EIA). WB: 0.25-0.5ug/ml|Human| IHC-P: 2-5ug/ml|Human| ICC/IF: 5ug/ml|Human| FC/FACS/FCM: 1-3ug/1x106 cells|Human| Direct ELISA: 0.1-0.5ug/ml|Human|. Researchers should empirically determine the suitability of the CYBB cybb for an application not listed in the data sheet. Researchers commonly develop new applications and it is an integral, important part of the investigative research process. It is sometimes possible for the material contained within the vial of "NOX2/gp91phox/CYBB, Polyclonal Antibody" to become dispersed throughout the inside of the vial, particularly around the seal of said vial, during shipment and storage. We always suggest centrifuging these vials to consolidate all of the liquid away from the lid and to the bottom of the vial prior to opening. Please be advised that certain products may require dry ice for shipping and that, if this is the case, an additional dry ice fee may also be required.Precautions
All products in the AAA Biotech catalog are strictly for research-use only, and are absolutely not suitable for use in any sort of medical, therapeutic, prophylactic, in-vivo, or diagnostic capacity. By purchasing a product from AAA Biotech, you are explicitly certifying that said products will be properly tested and used in line with industry standard. AAA Biotech and its authorized distribution partners reserve the right to refuse to fulfill any order if we have any indication that a purchaser may be intending to use a product outside of our accepted criteria.Disclaimer
Though we do strive to guarantee the information represented in this datasheet, AAA Biotech cannot be held responsible for any oversights or imprecisions. AAA Biotech reserves the right to adjust any aspect of this datasheet at any time and without notice. It is the responsibility of the customer to inform AAA Biotech of any product performance issues observed or experienced within 30 days of receipt of said product. To see additional details on this or any of our other policies, please see our Terms & Conditions page.Item has been added to Shopping Cart
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