|Validated Species:||Human, Mouse, Rat|
|Application:||Flow Cytometry, IHC, IHC-F, ICC, WB|
Data & Images
|Product Name||Anti-Ataxin 3 Antibody|
|Description||Rabbit IgG polyclonal antibody for Ataxin-3(ATXN3) detection. Tested with WB, IHC-P, IHC-F, ICC, FCM in Human;Mouse;Rat.|
|Cite This Product||Anti-Ataxin 3 Antibody (Boster Biological Technology, Pleasanton CA, USA, Catalog # PA1846)|
|Replacement Item||This antibody may replace the following items: sc-18480|sc-18483|sc-367680|sc-367744|sc-393193|sc-398114 from Santa Cruz Biotechnology.|
|Validated Species||Human, Mouse, Rat|
*This antibody is predicted to react with the above species based on antigen sequence similarities. Our Boster Guarantee covers the use of this product with the above species.
|Application||Flow Cytometry, IHC, IHC-F, ICC, WB
*Our Boster Guarantee covers the use of this product in the above tested applications.
**For positive and negative control design, consult "Tissue specificity" under Protein Target Info.
|Recommended Detection Systems||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), IHC(F) and ICC.
*Blocking peptide can be purchased at $50. Contact us for more information
**Boster also offers various secondary antibodies for Immunoflourescecne and IHC. Take advantage of the buy 1 primary antibody get 1 secondary antibody for free promotion for the entire year 2018!
|Immunogen||A synthetic peptide corresponding to a sequence at the N-terminus of human Ataxin 3(7-21aa EKQEGSLCAQHCLNN), identical to the related rat and mouse sequences.|
|Cross Reactivity||No cross reactivity with other proteins|
|Contents||Each vial contains 5mg BSA, 0.9mg NaCl, 0.2mg Na2HPO4, 0.05mg Thimerosal, 0.05mg NaN3.|
|Concentration||Add 0.2ml of distilled water will yield a concentration of 500ug/ml.|
|Storage||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.|
|Purification||Immunogen affinity purified.|
Protein Target Info (Source: Uniprot.org)
You can check the tissue specificity below for information on selecting positive and negative control.
|Molecular Weight||41781 MW|
|Protein Function||Deubiquitinating enzyme involved in protein homeostasis maintenance, transcription, cytoskeleton regulation, myogenesis and degradation of misfolded chaperone substrates. Binds long polyubiquitin chains and trims them, while it has weak or no activity against chains of 4 or less ubiquitins. Involved in degradation of misfolded chaperone substrates via its interaction with STUB1/CHIP: recruited to monoubiquitinated STUB1/CHIP, and restricts the length of ubiquitin chain attached to STUB1/CHIP substrates and preventing further chain extension. In response to misfolded substrate ubiquitination, mediates deubiquitination of monoubiquitinated STUB1/CHIP. Interacts with key regulators of transcription and represses transcription: acts as a histone- binding protein that regulates transcription. .|
|Sequence Similarities||Contains 1 Josephin domain.|
|Subcellular Localization||Nucleus matrix . Predominantly nuclear, but not exclusively, inner nuclear matrix.|
|Alternative Names||Ataxin-3;18.104.22.168;Machado-Joseph disease protein 1;Spinocerebellar ataxia type 3 protein;ATXN3;ATX3, MJD, MJD1, SCA3;|
|Research Areas|||epigenetics and nuclear signaling|chromatin modifying enzymes|ubiquitylation| neuroscience|neurology process|neurodegenerative disease| epigenetics and nuclear signaling|transcription|other factors|ubiquitin & ubiquitin like modifiers|deubiquitination||
Background for Ataxin-3
Dilution Ratios/Recommended Concentrations
At Boster we strive to provide the best Anti-Ataxin 3 Antibody by testing all applications on non-spiked tissues and cell lines to ensure that the affinity of the antibody is enough to react to the endogenouse level of the target protein. Read more about our QC panel here.
|Recommended dilution ratios are listed below:|
Immunohistochemistry(Paraffin-embedded Section), 0.5-1μg/ml, Human, Rat, Mouse, By Heat|
Western blot, 0.1-0.5μg/ml, Human, Rat, Mouse
Immunohistochemistry(Frozen Section), 0.5-1μg/ml, Human
Immunocytochemistry, 0.5-1μg/ml, Human
Flow Cytometry, 1-3μg/1x106 cells, Human
**Boster provides high sensitivity secondary antibody kits for Western blotting and IHC. For more info see Related Products below.
Anti-Ataxin 3 Antibody Images
Click the images to enlarge.
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: Rat Heart Tissue Lysate
Lane 3: Human Placenta Tissue Lysate
Lane 4: HELA Whole Cell Lysate
Lane 5: PANC Whole Cell Lysate
Lane 6: SMMC Whole Cell Lysate
Lane 7: COLO320 Whole Cell Lysate
Lane 8: MCF-7 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 # PA1846) 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.
Ataxin 3 was detected in paraffin-embedded section of human lung cancer tissues. 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 (PA1846) 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.
Overlay histogram showing A549 cells stained with PA1846 (Blue line).The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-Ataxin 3 Antibody (PA1846,1μg/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 (Red line) was also used as a control.
Overlay histogram showing MCF-7 cells stained with PA1846 (Blue line).The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-Ataxin 3 Antibody (PA1846,1μg/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 (Red line) was also used as a control.
Overlay histogram showing SiHa cells stained with PA1846 (Blue line).The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-Ataxin 3 Antibody (PA1846,1μg/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 (Red line) was also used as a control.
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
2. Post-translational cleavage: this can cause smaller bands and or multiple bands
3. Alternative splicing: the same gene can have alternative splicing patterns generating different size proteins, all with reactivities to the antibody.
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.
5. Multimers: Multimers are usually broken up in reducing conditions. However if the interactions between the multimers are strong, the band may appear higher.,