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Product Info Summary
| SKU: | A03119 |
|---|---|
| Size: | 100 μg/vial |
| Reactive Species: | Human |
| Host: | Rabbit |
| Application: | ELISA, Flow Cytometry, IF, IHC, ICC, WB |
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Product info
Product Name
Anti-NOX5 Antibody Picoband®
SKU/Catalog Number
A03119
Size
100 μg/vial
Form
Lyophilized
Description
Boster Bio Anti-NOX5 Antibody Picoband® catalog # A03119. Tested in ELISA, Flow Cytometry, IF, IHC, ICC, WB applications. This antibody reacts with Human. The brand Picoband indicates this is a premium antibody that guarantees superior quality, high affinity, and strong signals with minimal background in Western blot applications. Only our best-performing antibodies are designated as Picoband, ensuring unmatched performance.
Storage & Handling
Store at -20˚C for one year from date of receipt. After reconstitution, at 4˚C for one month. It can also be aliquotted and stored frozen at -20˚C for six months. Avoid repeated freeze-thaw cycles.
Cite This Product
Anti-NOX5 Antibody Picoband® (Boster Biological Technology, Pleasanton CA, USA, Catalog # A03119)
Host
Rabbit
Contents
Each vial contains 4 mg Trehalose, 0.9 mg NaCl and 0.2 mg Na2HPO4.
Clonality
Polyclonal
Isotype
Rabbit IgG
Immunogen
E.coli-derived human NOX5 recombinant protein (Position: A21-F765).
Cross-reactivity
No cross-reactivity with other proteins.
Reactive Species
A03119 is reactive to NOX5 in Human
Observed Molecular Weight
86 kDa
Calculated molecular weight
86.4 kDa
Background of NOX5
NOX5 (Nadph Oxidase 5), also known as NOX5A or NOX5B, is a protein which in humans is encoded by the NOX5 gene. The International Radiation Hybrid Mapping Consortium mapped the NOX5 gene to chromosome 15. NOX5 is a novel NADPH oxidase that generates superoxide and functions as an H+ channel in a Ca2+-dependent manner. It is found that, when heterologously expressed, NOX5 was quiescent in unstimulated cells. However, in response to elevations of the cytosolic Ca2+ concentration, it generated large amounts of superoxide. Using RT-PCR and Southern and Western blot analyses, NOX5 was identified as a flavin-containing Ca2+-dependent oxidase present in hairy leukemic cells (HC), but not normal B cells.
Antibody Validation
Boster validates all antibodies on WB, IHC, ICC, Immunofluorescence, and ELISA with known positive control and negative samples to ensure specificity and high affinity, including thorough antibody incubations.
Application & Images
Applications
A03119 is guaranteed for ELISA, Flow Cytometry, IF, IHC, ICC, WB Boster Guarantee
Assay Dilutions Recommendation
The recommendations below provide a starting point for assay optimization. The actual working concentration varies and should be decided by the user.
Western blot, 0.25-0.5μg/ml, Human
Immunohistochemistry (Paraffin-embedded Section), 2-5μg/ml, Human
Immunocytochemistry/Immunofluorescence, 5 μg/ml, Human
Flow Cytometry (Fixed), 1-3μg/1x106 cells, Human
ELISA, 0.1-0.5μg/ml, -
Positive Control
WB: human Jurkat whole cell, human Raji whole cell, human Hela whole cell, human 293T whole cell
IHC: human lung cancer tissue
ICC/IF: SiHa cell
FCM: Hela cell
Validation Images & Assay Conditions
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Western blot analysis of NOX5 using anti-NOX5 antibody (A03119).
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 30 ug of sample under reducing conditions.
Lane 1: human Jurkat whole cell lysates,
Lane 2: human Raji whole cell lysates,
Lane 3: human Hela whole cell lysates,
Lane 4: human 293T 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-NOX5 antigen affinity purified polyclonal antibody (Catalog # A03119) at 0.5 μg/mL overnight at 4°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 NOX5 at approximately 86 kDa. The expected band size for NOX5 is at 86 kDa.
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IHC analysis of NOX5 using anti-NOX5 antibody (A03119).
NOX5 was detected in a paraffin-embedded section of human lung cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 μg/ml rabbit anti-NOX5 Antibody (A03119) overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit (Catalog # SV0002) with DAB as the chromogen.
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IF analysis of NOX5 using anti-NOX5 antibody (A03119).
NOX5 was detected in an immunocytochemical section of SiHa cells. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent (AR0022) for 15 mins. The cells were blocked with 10% goat serum. And then incubated with 5 μg/mL rabbit anti-NOX5 Antibody (A03119) overnight at 4°C. DyLight®488 Conjugated Goat Anti-Rabbit IgG (BA1127) was used as secondary antibody at 1:500 dilution and incubated for 30 minutes at 37°C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used.
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Flow Cytometry analysis of Hela cells using anti-NOX5 antibody (A03119).
Overlay histogram showing Hela cells stained with A03119 (Blue line). The cells were fixed with 4% paraformaldehyde and blocked with 10% normal goat serum. And then incubated with rabbit anti-NOX5 Antibody (A03119, 1 μg/1x106 cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-rabbit IgG (BA1127, 5-10 μg/1x106 cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1 μg/1x106) used under the same conditions. Unlabelled sample without incubation with primary antibody and secondary antibody (Red line) was used as a blank control.
Specific Publications For Anti-NOX5 Antibody Picoband® (A03119)
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