Alpha tubulin Recombinant Rabbit Monoclonal Antibody [PSH02-94]
cat.: HA721913
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: PSH02-94
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
Images
HA721913_1.jpg 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.
HA721913_2.jpg 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.
HA721913_3.jpg 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.
HA721913_4.jpg 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.
HA721913_5.jpg 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.
HA721913_6.jpg 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.
HA721913_7.jpg 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.
HA721913_8.jpg 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.
HA721913_9.jpg 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).
HA721913_10.jpg 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
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.