Alpha tubulin Recombinant Rabbit Monoclonal Antibody [PSH02-95]
cat.: HA721914
| Product Type: | Recombinant Rabbit monoclonal IgG, primary antibodies |
|---|---|
| Species reactivity: | Human, Mouse, Rat |
| Applications: | WB, IHC-P, IF-Cell, FC, IP |
| Clonality: | Monoclonal |
| Clone number: | PSH02-95 |
| Form: | Liquid |
| Storage condition: | Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles. |
| Storage buffer: | PBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide. |
| Concentration: | 1ug/ul |
| Purification: | Protein A affinity purified. |
| Molecular weight: | Predicted band size: 50 kDa |
| Isotype: | IgG |
| Immunogen: | Synthetic peptide within Human Alpha-tubulin aa 402-451 / 451. |
| Positive control: | HeLa cell lysate, Daudi cell lysate, Jurkat cell lysate, A431 cell lysate, K-562 cell lysate, 293T cell lysate, NIH/3T3 cell lysate, PC-12 cell lysate, rat brain tissue lysate, mouse brain tissue lysate, mouse spleen tissue lysate, human tonsil tissue, rat brain tissue, Daudi, K-562, HeLa. |
| Subcellular location: | Cytoplasm, Cytoskeleton, Microtubule. |
| Recommended Dilutions:
WB IHC-P IF-Cell FC IP |
1:1,000-5,000 1:1,000 1:100 1:1,000 1-2μg/sample |
| Uniprot #: | SwissProt: P68363 Human | P68368 Mouse | Q5XIF6 Rat |
| Alternative names: | Alpha tubulin Alpha-tubulin Alpha-tubulin 1 ALS22 B ALPHA 1 bA408E5.3 H2 ALPHA Hum a tub1 Hum a tub2 LIS3 MGC171407 MGC55332 TBA4A_HUMAN Testis-specific alpha-tubulin TUBA1 TUBA1A tuba1l Tuba4a Tubulin alpha 1 chain Tubulin alpha Tubulin alpha-1 chain tubulin alpha-1B chain Tubulin alpha-4A chain Tubulin H2-alpha Tubulin, alpha 1 (testis specific) tubulin, alpha 1, like Tubulin, alpha 4a Tubulin, alpha, testis-specific Tubulin, alpha-1 |
Images
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Fig1:
Western blot analysis of Alpha tubulin on different lysates with Rabbit anti-Alpha tubulin antibody (HA721914) at 1/5,000 dilution. Lane 1: HeLa cell lysate Lane 2: Daudi cell lysate Lane 3: Jurkat cell lysate Lane 4: A431 cell lysate Lane 5: K-562 cell lysate Lane 6: 293T cell lysate Lane 7: NIH/3T3 cell lysate Lane 8: PC-12 cell lysate Lane 9: Rat brain tissue lysate Lane 10: Mouse brain tissue lysate Lane 11: Mouse spleen tissue lysate Cell lysates/proteins at 20 µg/Lane. Ttissue lysates/proteins at 40 µg/Lane. Predicted band size: 50 kDa Observed band size: 50 kDa Exposure time: 3 minutes; 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 (HA721914) at 1/5,000 dilution was used in 5% NFDM/TBST 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:
Immunofluorescence analysis of paraffin-embedded human tonsil tissue labeling Alpha tubulin with Rabbit anti-Alpha tubulin antibody (HA721914) at 1/1,000 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 10% negative goat serum for 1 hour at room temperature, washed with PBS, and then probed with the primary antibody (HA721914, green) at 1/1,000 dilution overnight at 4 ℃, washed with PBS. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. Nuclei were counterstained with DAPI (blue). |
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Fig3:
Immunohistochemical analysis of paraffin-embedded rat brain tissue with Rabbit anti-Alpha tubulin antibody (HA721914) at 1/1,000 dilution. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA721914) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX. |
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Fig4:
Immunocytochemistry analysis of Daudi cells labeling Alpha tubulin with Rabbit anti-Alpha tubulin antibody (HA721914) at 1/100 dilution. Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 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-Alpha tubulin antibody (HA721914) 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 (M1305-2, 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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Fig5:
Immunocytochemistry analysis of K-562 cells labeling Alpha tubulin with Rabbit anti-Alpha tubulin antibody (HA721914) at 1/100 dilution. Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 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-Alpha tubulin antibody (HA721914) 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 (M1305-2, 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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Fig6:
Flow cytometric analysis of HeLa cells labeling Alpha tubulin. Cells were fixed and permeabilized. Then stained with the primary antibody (HA721914, 1μg/mL) (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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Fig7:
Alpha tubulin was immunoprecipitated from 0.2 mg HeLa cell lysate with HA721914 at 2 µg/25 µl agarose. Western blot was performed from the immunoprecipitate using HA721914 at 1/1,000 dilution. Anti-Rabbit IgG for IP Nano-secondary antibody (NBI01H) at 1/5,000 dilution was used for 1 hour at room temperature. Lane 1: HeLa cell lysate (input) Lane 2: HA721914 IP in HeLa cell lysate Lane 3: Rabbit IgG instead of HA721914 in HeLa cell lysate Blocking/Dilution buffer: 5% NFDM/TBST Exposure time: 2 seconds; ECL: K1801 |
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.