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Product Info Summary
| SKU: | PB9423 |
|---|---|
| Size: | 100 μg/vial |
| Reactive Species: | Human, Rat |
| Host: | Rabbit |
| Application: | Flow Cytometry, IF, IHC, ICC, WB |
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Product info
Product Name
Anti-Ataxin 3/ATXN3 Antibody Picoband®
SKU/Catalog Number
PB9423
Size
100 μg/vial
Form
Lyophilized
Description
Boster Bio Anti-Ataxin 3/ATXN3 Antibody Picoband® catalog # PB9423. Tested in Flow Cytometry, IF, IHC, ICC, WB applications. This antibody reacts with Human, Rat. 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-Ataxin 3/ATXN3 Antibody Picoband® (Boster Biological Technology, Pleasanton CA, USA, Catalog # PB9423)
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 Ataxin 3, different from the related mouse and rat sequences by two amino acids.
Cross-reactivity
No cross-reactivity with other proteins
Reactive Species
PB9423 is reactive to ATXN3 in Human, Rat
Observed Molecular Weight
42 kDa
Calculated molecular weight
41.3 kDa
Background of ATXN3
ATXN3 (Ataxin 3), also known as AT3, MJD GENE, MJD1, SCA3 GENE, ATX3, JOS, Spinocerebellar ataxia-3, Machado-Joseph disease protein 1, is a protein that in humans is encoded by the ATXN3 gene. ATXN3 ranges in size from 360 to 374 amino acids. Using Northern blot analysis showed that ATXN3 mRNA was ubiquitously expressed in human tissues. They detected at least 4 ATXN3 transcripts of 1.4, 1.8, 4.5, and 7.5 kb and suggested that the different mRNA species probably result from differential splicing and polyadenylation. Machado-Joseph disease, also known as spinocerebellar ataxia-3, is an autosomal dominant neurologic disorder. The protein encoded by the ATXN3 gene contains (CAG)n repeats in the coding region, and the expansion of these repeats from the normal 13-36 to 68-79 is the cause of Machado-Joseph disease. There is an inverse correlation between the age of onset and CAG repeat numbers. Alternatively spliced transcript variants encoding different isoforms have been described for this gene. Ataxin-3 interacted with 2 human homologs of the yeast DNA repair protein RAD23, HHR23A (RAD23A) and HHR23B (RAD23B). Both normal and mutant ataxin-3 proteins interacted with the ubiquitin-like domain at the N terminus of the HHR23 proteins, which is a motif important for nucleotide excision repair. However, in HEK 293 cells, HHR23A was recruited to intranuclear inclusions formed by the mutant ataxin-3 through its interaction with ataxin-3.
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
PB9423 is guaranteed for 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, 0.1-0.5μg/ml, Human, Rat
Immunohistochemistry (Paraffin-embedded Section), 0.5-1μg/ml, Human
Immunocytochemistry/Immunofluorescence, 5μg/ml, Human
Flow Cytometry (Fixed), 1-3μg/1x106 cells, Human
Positive Control
WB: Rat Brain Tissue, COLO320 Whole Cell, HELA Whole Cell,
IHC: Human Lung Cancer tissue
ICC/IF: MCF-7 cell
FCM: A549 cell
Validation Images & Assay Conditions
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Western blot analysis of Ataxin 3 using anti-Ataxin 3 antibody (PB9423).
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 Brain Tissue Lysate
Lane 2: COLO320 Whole Cell Lysate
Lane 3: HELA Whole Cell Lysate
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-Ataxin 3 antigen affinity purified polyclonal antibody (Catalog # PB9423) 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 Ataxin 3 at approximately 42KD. The expected band size for Ataxin 3 is at 42KD.
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IHC analysis of Ataxin 3 using anti-Ataxin 3 antibody (PB9423).
Ataxin 3 was detected in paraffin-embedded section of Human Lung Cancer Tissue. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml rabbit anti-Ataxin 3 Antibody (PB9423) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC)(Catalog # SA1022) with DAB as the chromogen.
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Flow Cytometry analysis of A549 cells using anti-ATXN3 antibody (PB9423).
Overlay histogram showing A549 cells stained with PB9423 (Blue line). To facilitate intracellular staining, cells were fixed with 4% paraformaldehyde and permeabilized with permeabilization buffer. The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-ATXN3 Antibody (PB9423,1μg/1x106 cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-rabbit IgG (BA1127, 5-10μg/1x106 cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1μg/1x106) used under the same conditions. Unlabelled sample without incubation with primary antibody and secondary antibody (Red line) was used as a blank control.
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IF analysis of ATXN3 using anti-ATXN3 antibody (PB9423).
ATXN3 was detected in immunocytochemical section of MCF-7 cell. 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 5μg/mL rabbit anti-ATXN3 Antibody (PB9423) overnight at 4°C. DyLight®488 Conjugated Goat Anti-Rabbit IgG (BA1127) was used as secondary antibody at 1:100 dilution and incubated for 30 minutes at 37°C. Visualize using a fluorescence microscope and filter sets appropriate for the label used.
Specific Publications For Anti-Ataxin 3/ATXN3 Antibody Picoband® (PB9423)
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4 Customer Q&As for Anti-Ataxin 3/ATXN3 Antibody Picoband®
Question
We are currently using anti-Ataxin 3/ATXN3 antibody PB9423 for human tissue, and we are well pleased with the WB results. The species of reactivity given in the datasheet says human, rat. Is it likely that the antibody can work on dog tissues as well?
Verified Customer
Verified customer
Asked: 2019-09-25
Answer
The anti-Ataxin 3/ATXN3 antibody (PB9423) has not been tested for cross reactivity specifically with dog 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 dog you can get your next antibody for free. Please contact me if I can help you with anything.
Boster Scientific Support
Answered: 2019-09-25
Question
I see that the anti-Ataxin 3/ATXN3 antibody PB9423 works with WB, what is the protocol used to produce the result images on the product page?
Verified Customer
Verified customer
Asked: 2019-06-12
Answer
You can find protocols for WB on the "support/technical resources" section of our navigation menu. If you have any further questions, please send an email to support@bosterbio.com
Boster Scientific Support
Answered: 2019-06-12
Question
Thank you for helping with my inquiry over the phone. Here are the WB image, lot number and protocol we used for corpus callosum using anti-Ataxin 3/ATXN3 antibody PB9423. Let me know if you need anything else.
R. Anderson
Verified customer
Asked: 2018-02-19
Answer
Thank you for 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: 2018-02-19
Question
I was wanting to use to test anti-Ataxin 3/ATXN3 antibody PB9423 on rat corpus callosum for research purposes, then I may be interested in using anti-Ataxin 3/ATXN3 antibody PB9423 for diagnostic purposes as well. Is the antibody suitable for diagnostic purposes?
J. Li
Verified customer
Asked: 2013-03-22
Answer
The products we sell, including anti-Ataxin 3/ATXN3 antibody PB9423, are only intended for research use. They would not be suitable for use in diagnostic work. If you have the means to develop a product into diagnostic use, and are interested in collaborating with us and develop our product into an IVD product, please contact us for more discussions.
Boster Scientific Support
Answered: 2013-03-22
