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Product Info Summary
| SKU: | PA1947 |
|---|---|
| Size: | 100 μg/vial |
| Reactive Species: | Human |
| Host: | Rabbit |
| Application: | IF, IHC, ICC, WB |
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Product info
Product Name
Anti-NADPH oxidase 5 NOX5 Antibody Picoband®
SKU/Catalog Number
PA1947
BA2819 is an alternative SKU for this antibody, used in previous lots.
Size
100 μg/vial
Form
Lyophilized
Description
Boster Bio Anti-NADPH oxidase 5 NOX5 Antibody catalog # PA1947. Tested in IHC, ICC/IF, 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-NADPH oxidase 5 NOX5 Antibody Picoband® (Boster Biological Technology, Pleasanton CA, USA, Catalog # PA1947)
Host
Rabbit
Contents
Each vial contains 4 mg Trehalose, 0.9 mg NaCl and 0.2 mg Na2HPO4.
Clonality
Polyclonal
Isotype
Rabbit IgG
Immunogen
A synthetic peptide corresponding to a sequence at the C-terminus of human NOX5.
Cross-reactivity
No cross-reactivity with other proteins
Reactive Species
PA1947 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. Banfi et al. (2001) 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, Kamiguti et al. (2005) identified NOX5 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
PA1947 is guaranteed for 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, 0.1-0.5μg/ml, Human
Immunohistochemistry (Paraffin-embedded Section), 2-5μg/ml, Human
Immunocytochemistry/Immunofluorescence, 5 μg/ml, Human
Positive Control
WB: human Jurkat whole cell, human Raji whole cell, human Hela whole cell, human 293T whole cell
IHC: human prostatic cancer tissue
ICC/IF: U2OS cell
Validation Images & Assay Conditions
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Western blot analysis of NOX5 using anti-NOX5 antibody (PA1947).
Electrophoresis was performed on a 10% SDS-PAGE gel at 80V (Stacking gel) / 120V (Resolving gel) for 2 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 150 mA 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 (PA1947) 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 (Catalog # BA1054) at a dilution of 1:5000 for 1.5 hour at RT. The signal is developed using an ECL Plus Western Blotting Substrate (Catalog # AR1196-200) 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 (PA1947).
NOX5 was detected in a paraffin-embedded section of human prostatic 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 (PA1947) 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 (PA1947).
NOX5 was detected in an immunocytochemical section of U2OS 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 (PA1947) overnight at 4°C. Fluoro488 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.
Specific Publications For Anti-NADPH oxidase 5 NOX5 Antibody Picoband® (PA1947)
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3 Customer Q&As for Anti-NADPH oxidase 5 NOX5 Antibody Picoband®
Question
We are currently using anti-NOX5 antibody PA1947 for human tissue, and we are satisfied with the IHC results. The species of reactivity given in the datasheet says human. Is it true that the antibody can work on bovine tissues as well?
Verified Customer
Verified customer
Asked: 2019-08-15
Answer
The anti-NOX5 antibody (PA1947) has not been validated for cross reactivity specifically with bovine tissues, though there is a good chance of cross reactivity. We have an innovator award program that if you test this antibody and show it works in bovine you can get your next antibody for free. Please contact me if I can help you with anything.
Boster Scientific Support
Answered: 2019-08-15
Question
Would anti-NOX5 antibody PA1947 work on canine ICC with spleen testis?
Verified Customer
Verified customer
Asked: 2019-07-18
Answer
Our lab technicians have not tested anti-NOX5 antibody PA1947 on canine. You can run a BLAST between canine and the immunogen sequence of anti-NOX5 antibody PA1947 to see if they may cross-react. If the sequence homology is close, then you can perform a pilot test. Keep in mind that since we have not validated canine samples, this use of the antibody is not covered by our guarantee. However we have an innovator award program that if you test this antibody and show it works in canine spleen testis in ICC, you can get your next antibody for free.
Boster Scientific Support
Answered: 2019-07-18
Question
We appreciate helping with my inquiry over the phone. Here are the WB image, lot number and protocol we used for kidney epithelium spleen using anti-NOX5 antibody PA1947. Let me know if you need anything else.
L. Rodriguez
Verified customer
Asked: 2014-01-09
Answer
We appreciate the data. You have provided everything we needed. Our lab team are working to resolve your inquiry as quickly as possible, and we appreciate your patience and understanding! Please let me know if there is anything you need in the meantime.
Boster Scientific Support
Answered: 2014-01-09
