Product Type: | Recombinant Rabbit monoclonal IgG, primary antibodies |
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Species reactivity: | Human, Mouse, Rat |
Applications: | WB, IHC-P, IF-Cell, FC, IP |
Clonality: | Monoclonal |
Clone number: | PSH02-94 |
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: 55 kDa |
Isotype: | IgG |
Immunogen: | Synthetic peptide within Human Alpha-tubulin aa 402-451 / 451. |
Positive control: | HeLa cell lysate, PC-12 cell lysate, NIH/3T3 cell lysate, Daudi cell lysate, Jurkat cell lysate, A431 cell lysate, K-562 cell lysate, rat kidney tissue lysate, rat heart tissue lysate, rat liver tissue lysate, human tonsil tissue, rat brain tissue, Daudi, K-562, HeLa, NIH/3T3. |
Subcellular location: | Cytoplasm, Cytoskeleton, Microtubule. |
Recommended Dilutions:
WB IHC-P IF-Cell FC IP |
1:1,000-1:5,000 1:1,000 1:250-1:500 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 |
Fig1:
Western blot analysis of Alpha tubulin on different lysates with Rabbit anti-Alpha tubulin antibody (HA721913) at 1/5,000 dilution. Lane 1: HeLa cell lysate Lane 2: PC-12 cell lysate Lane 3: NIH/3T3 cell lysate Lane 4: Daudi cell lysate Lane 5: Jurkat cell lysate Lane 6: A431 cell lysate Lane 7: K-562 cell lysate Lysates/proteins at 20 µg/Lane. Predicted band size: 50 kDa Observed band size: 50 kDa Exposure time: 1 minutes 2 seconds; 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 (HA721913) 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:
Western blot analysis of Alpha tubulin on different lysates with Rabbit anti-Alpha tubulin antibody (HA721913) at 1/1,000 dilution. Lane 1: Rat kidney tissue lysate (40 µg/Lane) Lane 2: Rat heart tissue lysate (40 µg/Lane) Lane 2: Rat liver tissue lysate (40 µg/Lane) Predicted band size: 50 kDa Observed band size: 50 kDa Exposure time: 30 seconds; 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 (HA721913) at 1/1,000 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. |
Fig3:
Immunohistochemical analysis of paraffin-embedded human tonsil tissue with Rabbit anti-Alpha tubulin antibody (HA721913) 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 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA721913) 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:
Immunohistochemical analysis of paraffin-embedded rat brain tissue with Rabbit anti-Alpha tubulin antibody (HA721913) 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 (HA721913) 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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Fig5:
Immunocytochemistry analysis of Daudi cells labeling Alpha tubulin with Rabbit anti-Alpha tubulin antibody (HA721913) at 1/500 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 (HA721913) at 1/500 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. |
Fig6:
Immunocytochemistry analysis of K-562 cells labeling Alpha tubulin with Rabbit anti-Alpha tubulin antibody (HA721913) at 1/250 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 (HA721913) at 1/250 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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Fig7:
Immunocytochemistry analysis of HeLa cells labeling Alpha tubulin with Rabbit anti-Alpha tubulin antibody (HA721913) at 1/500 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 (HA721913) at 1/500 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. |
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Fig8:
Immunocytochemistry analysis of NIH/3T3 cells labeling Alpha tubulin with Rabbit anti-Alpha tubulin antibody (HA721913) at 1/500 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 (HA721913) at 1/500 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. |
Fig9:
Flow cytometric analysis of HeLa cells labeling Alpha tubulin. Cells were fixed and permeabilized. Then stained with the primary antibody (HA721913, 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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Fig10:
Alpha tubulin was immunoprecipitated from 0.2 mg HeLa cell lysate with HA721913 at 2 µg/25 µl agarose. Western blot was performed from the immunoprecipitate using HA721913 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: HA721913 IP in HeLa cell lysate Lane 3: Rabbit IgG instead of HA721913 in HeLa cell lysate Blocking/Dilution buffer: 5% NFDM/TBST Exposure time: 2 seconds; ECL: K1801 |