|Reactivity||Human, Mouse, Rat|
|Product Name||Anti-NOX2/gp91phox/CYBB Antibody|
|Storage & Handling||At -20°C for one year. After reconstitution, at 4°C for one month. It can also be aliquotted and stored frozen at -20°C for a longer time.Avoid repeated freezing and thawing.|
|Description||Rabbit IgG polyclonal antibody for Cytochrome b-245 heavy chain(CYBB) detection. Tested with WB, IHC-P in Human;Mouse;Rat.|
|Cite This Product||Anti-NOX2/gp91phox/CYBB Antibody (Boster Biological Technology, Pleasanton CA, USA, Catalog # PA1667)|
|Contents/Buffer||Each vial contains 5mg BSA, 0.9mg NaCl, 0.2mg Na2HPO4, 0.05mg Thimerosal, 0.05mg NaN3.|
|Immunogen||A synthetic peptide corresponding to a sequence in the middle region of human NOX2(442-459aa YWLCRDTHAFEWFADLLQ), identical to the related rat and mouse sequences.|
|Reactivity||Human, Mouse, Rat|
Assay Dilutions Overview
Western blot, 0.1-0.5μg/ml, Human, Rat, Mouse
Immunohistochemistry(Paraffin-embedded Section), 0.5-1μg/ml, Human, Rat, Mouse, By Heat
Boster's Secondary Antibodies And IHC, WB Kits
The following reagents are used to generate the images below.Boster recommends Enhanced Chemiluminescent Kit with anti-Rabbit IgG (EK1002) for Western blot, and HRP Conjugated anti-Rabbit IgG Super Vision Assay Kit (SV0002-1) for IHC(P).
Images And Assay Conditions
Anti-NOX2/gp91phox antibody, PA1667, Western blotting
Lane 1: HELA Cell Lysate
Lane 2: JURKAT Cell Lysate
Lane 3: MCF-7 Cell Lysate
Lane 4: SMMC Cell Lysate
Anti-NOX2/gp91phox antibody, PA1667, IHC(P)
IHC(P): Human Intestinal Cancer Tissue
Figure 3. Western blot analysis of NOX2 using anti- NOX2 antibody (PA1667).
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: rat thymus tissue lysates,
Lane 2: rat brain tissue 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 antigen affinity purified polyclonal antibody (Catalog # PA1667) at 0.5 Î¼g/mL overnight at 4Â°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 1:10000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # EK1002) with Tanon 5200 system. A specific band was detected for NOX2 at approximately 65KD. The expected band size for NOX2 is at 65KD.
Protein Target Info (Source: Uniprot.org)
|Protein Name||Cytochrome b-245 heavy chain|
|Tissue Specificity||Detected in neutrophils (at protein level). .|
|Alternative Names||Cytochrome b-245 heavy chain;1.-.-.-;CGD91-phox;Cytochrome b(558) subunit beta;Cytochrome b558 subunit beta;Heme-binding membrane glycoprotein gp91phox;NADPH oxidase 2;Neutrophil cytochrome b 91 kDa polypeptide;Superoxide-generating NADPH oxidase heavy chain subunit;gp91-1;gp91-phox;p22 phagocyte B-cytochrome;CYBB;NOX2;|
|Subcellular Localization||Cell membrane; Multi-pass membrane protein.|
|Molecular Weight||65336 MW|
*if product is indicated to react with multiple species, protein info is based on the human gene.
|Protein Function||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.|
|Research Areas||Human, Mouse, Rat
*You can search these to find other products in these research areas.
|Background||NOX2(NADPH OXIDASE 2), also called CYBB(CYTOCHROME b(-245), BETA SUBUNIT), p91-PHOX or GP91-1, is a human gene encoding a glycoprotein.NOX2 is an essential component of phagocytic NADPH-oxidase, a membrane-bound enzyme complex that generates large quantities of microbicidal superoxide and other oxidants upon activation. It is mapped on Xp11.4. NOX2 is a heterodimer composed of an alpha chain of relative molecular mass 23 kD and a beta chain of 76 to 82 kD. NOX2 assembled on DC phagosomes in a gp91-phox subunit-dependent manner, and that reactive oxygen species were produced in a more sustained manner in immature DC phagosomes than in macrophage phagosomes. As a major player in innate immune responses in neutrophils, NOX2 is also involved in adaptive immunity through its activity in DCs. In heart cells, physiologic stretch rapidly activates reduced-form NOX2 to produce reactive oxygen species(ROS) in a process dependent on microtubules(X-ROS signaling).|
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Q: Do you offer BSA-free antibodies? Keyword: Bovine serum albumin, carrier protein, conjugationA: Yes, please contact us at email@example.com for more information about BSA-free antibodies and availability. The new BSA-free formula uses trehalose as a replacement to BSA. We have tested many alternative chemicals and found that trehalose protects the antibodies the best.
Q: Is your western blot protocol provided from the website applicable for all your antibodies? Keyword: applications, WBA: The protocol is applicable for all our antibodies in WB, the NC Membrane(0.45μm or 0.22μm) and transfer time(70 mins or 50 mins) depends on the protein molecular weight, details can be found in included protocol.
Q: Can I conjugate markers to this antibody? Can I link custom conjugates to this antibody? Keyword: conjugationA: The antibody is stored with BSA and cannot be conjugated with markers. Carrier free antibodies are available upon request. Please contact firstname.lastname@example.org
Q: What should I use for negative control?A: Please contact us for negative control suggestions. You can also check expression databases such as genecards, uniprot etc. Due to logistic reasons, we do not sell serum or lysates that we use internally for positive or negative control.
Q: Where can I find troubleshooting information? What should I do if I have unexpected bands, high background, no signal, weak signalA: You can find Boster's troubleshoot guides under tech support tab. Please contact us for further assistance on troubleshooting your experiment.
Q: What is the immunogen sequence of this antibody? Is this antibody polyclonal or monoclonal?A: You can find the immunogen sequence under "
Q: What is the expected band size? Why is it different than the observed band size?A: The expected band size is predicted on the size of the protein. The actual band size may be affected by a few other factors including but not limited to:<br>1. Post-translational modification:phosphorylation, methylation, glycosylation etc. These modifications prevent SDS molecules from binding to the target protein and thus make the band size appear larger than expected<br>2. Post-translational cleavage: this can cause smaller bands and or multiple bands <br><br>3. Alternative splicing: the same gene can have alternative splicing patterns generating different size proteins, all with reactivities to the antibody. <br><br>4. Amino Acid R chain charge: SDS binds to positive charges. The different size and charge of the Amino Acid side chains can affect the amount of SDS binding and thus affect the observed band size.<br>5. Multimers: Multimers are usually broken up in reducing conditions. However if the interactions between the multimers are strong, the band may appear higher., <br>
Q: What is the suggested dilution ratio for Western Blot (WB), Immunohistochemistry (IHC) and or ELISA standards? What is the optimal pH for the sample?A: Check the datasheet for the product for details on dilution ratios for different experiments. You can find the datasheet button on the right side of the product page.
Q: What is the protocol you used for your Western blotting (WB) and Immunohistochemistry (IHC)?A: Check our protocols under the tech support tab.
Q: What are some alternative names that could be used to describe this product?A: Some common names include but are not limited to gp91 phox antibody, gp91phox antibody, gp91-phox antibody, nox2 antibody