Ataxin 3 Recombinant Rabbit Monoclonal Antibody [PSH20-73]
cat.: HA751768
| Product Type: | Recombinant Rabbit monoclonal IgG, primary antibodies |
|---|---|
| Species reactivity: | Human |
| Applications: | WB, IF-Cell, FC, IP |
| Clonality: | Monoclonal |
| Clone number: | PSH20-73 |
| Form: | Liquid |
| Storage condition: | Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles. |
| Storage buffer: | 1*PBS (pH7.4). |
| Concentration: | 1ug/ul |
| Purification: | Protein A affinity purified. |
| Molecular weight: | Predicted band size: 41 kDa |
| Isotype: | IgG |
| Immunogen: | Recombinant protein within human Ataxin 3 aa 1-361. |
| Positive control: | U-87 MG cell lysate, HeLa cell lysate, HEK-293 cell lysate, SH-SY5Y cell lysate, A549 cell lysate, U-87 MG. |
| Subcellular location: | Nucleus matrix, Nucleus, Lysosome membrane. |
| Recommended Dilutions:
WB IF-Cell FC IP |
1:5,000 1:100 1:1,000 1-2μg/sample |
| Uniprot #: | SwissProt: P54252 Human |
| Alternative names: | AT3 Ataxin 3 ataxin 3 variant h ataxin 3 variant m ataxin 3 variant ref 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 |
Images
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Fig1:
Western blot analysis of Ataxin 3 on different lysates with Rabbit anti-Ataxin 3 antibody (HA751768) at 1/5,000 dilution. Lane 1: U-87 MG cell lysate (20 µg/Lane) Lane 2: HeLa cell lysate (20 µg/Lane) Lane 3: HEK-293 cell lysate (20 µg/Lane) Lane 4: SH-SY5Y cell lysate (20 µg/Lane) Lane 5: A549 cell lysate (20 µg/Lane) Lysates/proteins at 20 µg/Lane. Predicted band size: 41 kDa Observed band size: 45 kDa Exposure time: 20 seconds; ECL: K1801; 4-20% SDS-PAGE gel. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA751768) at 1/5,000 dilution was used in primary antibody dilution (K1803) at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature. |
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Fig2:
Immunocytochemistry analysis of U-87 MG cells labeling Ataxin 3 with Rabbit anti-Ataxin 3 antibody (HA751768) at 1/100 dilution. Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-Ataxin 3 antibody (HA751768) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI. Beta tubulin (HA601187, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution. |
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Fig3:
Flow cytometric analysis of U-87 MG cells labeling Ataxin 3. Cells were fixed and permeabilized. Then stained with the primary antibody (HA751768, 1/1,000) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black). |
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Fig4:
Ataxin 3 was immunoprecipitated from 0.2 mg HEK-293 cell lysate with HA751768 at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using HA751768 at 1/5,000 dilution. HRP Conjugated Anti-Rabbit IgG for IP Nano-secondary antibody at 1/5,000 dilution was used for 1 hour at room temperature. Lane 1: HEK-293 cell lysate (input) Lane 2: HA751768 IP in HEK-293 cell lysate Lane 3: Rabbit IgG instead of HA751768 in HEK-293 cell lysate Blocking/Dilution buffer: primary antibody dilution (K1803) Exposure time: 1 30 seconds; ECL: K1801 |
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.