Anti-Ataxin 3 Antibody | MyBioSource MBS178269 product information

product summary

company name :

MyBioSource

product type :

antibody

product name :

Anti-Ataxin 3 Antibody

catalog :

MBS178269

quantity :

0.1 mg

price :

315 USD

clonality :

polyclonal

host :

unidentified

conjugate :

nonconjugated

reactivity :

human, rat

application :

western blot, immunohistochemistry, immunocytochemistry, flow cytometry, immunohistochemistry - paraffin section, immunohistochemistry - frozen section

more info or order :

MyBioSource product webpage

image

image 1 :

Figure 1. 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 at 0.5 ug/mL overnight at 4 degree 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 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.

image 2 :

Figure 2. IHC analysis of Ataxin 3 using anti-Ataxin 3 antibody. 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 1ug/ml rabbit anti-Ataxin 3 Antibody (PB9423) overnight at 4 degree C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) with DAB as the chromogen.

product information

catalog number :

MBS178269

products type :

Antibody

products full name :

Anti-Ataxin 3 Antibody

products short name :

[Ataxin 3]

products name syn :

[Ataxin-3; AT3; Ataxin 3; Ataxin-3; ATX3; ATX3_HUMAN; ATXN3; EC 3.4.22.; JOS; Josephin; Machado Joseph disease (spinocerebellar ataxia 3, olivopontocerebellar ataxia 3, autosomal dominant, ataxin 3); Machado Joseph disease; Machado Joseph disease protein 1; Machado-Joseph disease protein 1; Machado-Joseph disease protein 1 homolog; MJD; MJD gene; MJD1; Olivopontocerebellar ataxia 3; OTTHUMP00000221583; OTTHUMP00000221585; OTTHUMP00000221586; OTTHUMP00000221587; OTTHUMP00000231995; OTTHUMP00000231997; Rsca3; SCA3; SCA3 gene; Spinocerebellar ataxia type 3 protein; ataxin 3]

other names :

[ataxin-3 isoform ad; Ataxin-3; ataxin-3; ataxin 3; Machado-Joseph disease protein 1; Spinocerebellar ataxia type 3 protein]

other gene names :

[ATXN3; ATXN3; AT3; JOS; MJD; ATX3; MJD1; SCA3; ATX3; MJD; MJD1; SCA3]

uniprot entry name :

ATX3_HUMAN

clonality :

Polyclonal

host :

unknown

reactivity :

Human, Rat

sequence length :

346

specificity :

No cross reactivity with other proteins.

purity :

Immunogen Affinity Purified

form :

Lyophilized. Each vial contains 5mg BSA, 0.9mg NaCl, 0.2mg Na2HPO4, 0.05mg NaN3.

storage stability :

At -20 degree C for one year. After reconstitution, at 4 degree C for one month. It can also be aliquoted and stored frozen at -20 degree C for a longer time. Avoid repeated freezing and thawing.

tested application :

Western Blot (WB), Immunohistochemistry (IHC) Paraffin/Frozen, Immunocytochemistry (ICC), Flow Cytometry (FC)

app notes :

Western Blot: Concentration: 0.1-0.5ug/ml. Tested Species: Human, Rat. Antigen Retrieval: N/A. Immunohistochemistry - Paraffin: Concentration: 0.5-1ug/ml. Tested Species: Human. Antigen Retrieval: By Heat. Immunohistochemistry - Frozen: Concentration: 0.5-1ug/ml. Tested Species: Human. Antigen Retrieval: N/A. Immunocytochemistry: Concentration: 0.5-1ug/ml. Tested Species: Human. Antigen Retrieval: N/A. Flow Cytometry: 1-3ug/1x10 6 cells. Tested Species: Human. Antigen Retrieval: N/A. Tested Species: In-house tested species with positive results. By Heat: Boiling the paraffin sections in 10mM citrate buffer, pH 6.0, for 20 mins is required for the staining of formalin/paraffin sections. . Other applications have not been tested. Optimal dilutions should be determined by end users.

image1 heading :

Western Blot (WB)

image2 heading :

Immunohistochemistry (IHC)

image3 heading :

Flow Cytometry (FC/FACS)

image4 heading :

Flow Cytometry (FC/FACS)

image4 description :

Figure 4. Flow Cytometry analysis of SiHa cells using anti-ATXN3 antibody (PB9423).Overlay histogram showing SiHa cells stained with PB9423 (Blue line).The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-ATXN3 Antibody (1ug/1x106 cells) for 30 min at 20 degree C. DyLight 488 conjugated goat anti-rabbit IgG (5-10ug/1x106 cells) was used as secondary antibody for 30 minutes at 20 degree C. Isotype control antibody (Green line) was rabbit IgG (1ug/1x106) used under the same conditions. Unlabelled sample (Red line) was also used as a control.

image5 heading :

Immunofluorescence (IF)

image5 description :

Figure 5. IF analysis of ATXN3 using anti- ATXN3 antibody (PB9423). ATXN3 was detected in immunocytochemical section of U20S cell. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent for 15 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2ug/mL rabbit anti- ATXN3 Antibody overnight at 4 degree C. DyLight 488 Conjugated Goat Anti-Rabbit IgG was used as secondary antibody at 1:100 dilution and incubated for 30 minutes at 37 degree C. Visualize using a fluorescence microscope and filter sets appropriate for the label used.

other info1 :

Immunogen: A synthetic peptide corresponding to a sequence at the C-terminus of human Ataxin 3 (226-254aa EEDLQRALALSRQEIDMEDEEADLRRAIQ), different from the related mouse and rat sequences by two amino acids. Ig Type: Rabbit IgG

other info2 :

Reconstitution: Add 0.2ml of distilled water will yield a concentration of 500ug/ml.

products description :

Description: Rabbit IgG polyclonal antibody for Ataxin-3(ATXN3) detection. Tested with WB, IHC-P in Human;Rat. Background: 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.

products references :

1. Burnett, B., Li, F., Pittman, R. N. The polyglutamine neurodegenerative protein ataxin-3 binds polyubiquitylated proteins and has ubiquitin protease activity. Hum. Molec. Genet. 12: 3195-3205, 2003. 2. Paulson, H. L., Das, S. S., Crino, P. B., Perez, M. K., Patel, S. C., Gotsdiner, D., Fischbeck, K. H., Pittman, R. N.Machado-Joseph disease gene product is a cytoplasmic protein widely expressed in brain.Ann. Neurol. 41: 453-462, 1997.

ncbi gi num :

189163493

ncbi acc num :

NP_001121168.1

ncbi gb acc num :

NM_001127696.1

uniprot acc num :

P54252

ncbi pathways :

Protein Processing In Endoplasmic Reticulum Pathway (167325); Protein Processing In Endoplasmic Reticulum Pathway (167190)

ncbi summary :

Machado-Joseph disease, also known as spinocerebellar ataxia-3, is an autosomal dominant neurologic disorder. The protein encoded by this gene contains (CAG)n repeats in the coding region, and the expansion of these repeats from the normal 13-36 to 68-79 is one cause of Machado-Joseph disease. There is a negative correlation between the age of onset and CAG repeat numbers. Alternatively spliced transcript variants encoding different isoforms have been described for this gene. [provided by RefSeq, Sep 2009]

uniprot summary :

ataxin-3: 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. Defects in ATXN3 are the cause of spinocerebellar ataxia type 3 (SCA3); also known as Machado-Joseph disease (MJD). Spinocerebellar ataxia is a clinically and genetically heterogeneous group of cerebellar disorders. Patients show progressive incoordination of gait and often poor coordination of hands, speech and eye movements, due to degeneration of the cerebellum with variable involvement of the brainstem and spinal cord. SCA3 belongs to the autosomal dominant cerebellar ataxias type I (ADCA I) which are characterized by cerebellar ataxia in combination with additional clinical features like optic atrophy, ophthalmoplegia, bulbar and extrapyramidal signs, peripheral neuropathy and dementia. The molecular defect in SCA3 is the a CAG repeat expansion in ATXN3 coding region. Longer expansions result in earlier onset and more severe clinical manifestations of the disease. 3 isoforms of the human protein are produced by alternative splicing. Protein type: Ubiquitin-specific protease; DNA repair, damage; Transcription regulation; Protease; EC 3.4.19.12. Chromosomal Location of Human Ortholog: 14q21. Cellular Component: cytoplasm; cytosol; endoplasmic reticulum membrane; mitochondrial matrix; mitochondrial membrane; nuclear inclusion body; nuclear matrix; nucleoplasm; nucleus. Molecular Function: ATPase binding; histone deacetylase activity; identical protein binding; omega peptidase activity; protein binding; ubiquitin protein ligase binding; ubiquitin-specific protease activity. Biological Process: actin cytoskeleton organization and biogenesis; intermediate filament cytoskeleton organization and biogenesis; microtubule cytoskeleton organization and biogenesis; misfolded or incompletely synthesized protein catabolic process; nervous system development; nucleotide-excision repair; proteasomal ubiquitin-dependent protein catabolic process; protein deubiquitination; regulation of transcription, DNA-dependent; synaptic transmission; transcription, DNA-dependent; ubiquitin-dependent protein catabolic process. Disease: Machado-joseph Disease

size1 :

0.1 mg

price1 :

315 USD

more info or order :

MyBioSource product webpage