alpha Tubulin Rabbit Polyclonal Antibody
cat.: ER130905
Product Type: Rabbit polyclonal IgG, primary antibodies
Species reactivity: Human, Mouse, Rat, Zebrafish
Applications: WB, IHC-P, FC, IF-Cell, IF-Tissue
Clonality: Polyclonal
Form: Liquid
Storage condition: Store at +4℃ after thawing. Aliquot store at -20℃ or -80℃. Avoid repeated freeze / thaw cycles.
Storage buffer: 1*PBS (pH7.4), 0.2% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
Concentration: 1ug/ul
Purification: Immunogen affinity purified.
Molecular weight: Predicted band size: 50 kDa
Isotype: IgG
Immunogen: Synthetic peptide within Human Alpha-tubulin aa 402-451 / 451.
Positive control: NIH/3T3 cell lysate, HepG2 cell lysate, PC-12 cell lysate, Hela cell lysate, HepG2, Hela, Neuro-2a, C6, human stomach tissue, human tonsil tissue, mouse pancreas tissue, rat large intestine tissue.
Subcellular location: Cytoplasm, Cytoskeleton, Microtubule.
Recommended Dilutions:
  WB
  IHC-P
  FC
  IF-Cell
  IF-Tissue
1:1,000-1:10,000
1:200-1:500
1:50-1:100
1:200-1:500
1:200
Uniprot #: SwissProt: P68366 Human | P68368 Mouse | Q5XIF6 Rat
Alternative names: TUBA1 TUBA1A TUBA2 TUBA3 TUBA3C TUBA4A Tubulin, alpha 1a Tubulin, alpha 3c Tubulin, alpha 4a
Images
ER130905_1.jpg Fig1: Western blot analysis of alpha Tubulin on different lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (ER130905, 1/10,000) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:200,000 dilution was used for 1 hour at room temperature.
Positive control:
Lane 1: NIH/3T3 cell lysate
Lane 2: HepG2 cell lysate
Lane 3: PC-12 cell lysate
Lane 4: Hela cell lysate
ER130905_2.jpg Fig2: Western blot analysis of alpha Tubulin on different lysates with Rabbit anti-alpha Tubulin antibody (ER130905) at different dilutions.

Lane 1/2: NIH3T3 cell lysate at 1/1,000 and 1/5,000 dilution
Lane 3/4: HepG2 cell lysate at 1/1,000 and 1/5,000 dilution
Lane 5/6: PC-12 cell lysate at 1/1,000 and 1/5,000 dilution
Lane 7/8: Hela cell lysate at 1/1,000 and 1/5,000 dilution

Lysates/proteins at 10 µg/Lane.

Predicted band size: 50 kDa
Observed band size: 50 kDa

Exposure time: 1 minute;

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 (ER130905) at different dilutions were used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:100,000 dilution was used for 1 hour at room temperature.
ER130905_3.jpg Fig3: ICC staining of alpha Tubulin in HepG2 cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 10% negative goat serum for 15 minutes at room temperature. Cells were probed with the primary antibody (ER130905, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 conjugate-Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
ER130905_4.jpg Fig4: ICC staining of alpha Tubulin in Hela cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 10% negative goat serum for 15 minutes at room temperature. Cells were probed with the primary antibody (ER130905, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 conjugate-Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
ER130905_5.jpg Fig5: Immunocytochemistry analysis of Neuro-2a cells labeling alpha Tubulin with Rabbit anti-alpha Tubulin antibody (ER130905) 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 (ER130905) 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.
ER130905_6.jpg Fig6: Immunocytochemistry analysis of C6 cells labeling alpha Tubulin with Rabbit anti-alpha Tubulin antibody (ER130905) 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 (ER130905) 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.
ER130905_7.jpg Fig7: Immunofluorescence staining of paraffin-embedded human stomach tissue using anti-alpha Tubulin antibody. 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 ER130905 at 1/50 dilution for 10 hours at 4℃ and detected using Alexa Fluor® 594 conjugate-Goat anti-Rabbit IgG (H+L) Secondary Antibody at a dilution of 1:500 for 1 hour at room temperature.
ER130905_8.jpg Fig8: Immunohistochemical analysis of paraffin-embedded human tonsil tissue using anti-alpha Tubulin antibody. 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 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ER130905, 1/50) for 30 minutes 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.
ER130905_9.jpg Fig9: Immunohistochemical analysis of paraffin-embedded mouse pancreas tissue using anti-alpha Tubulin antibody. 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 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ER130905, 1/50) for 30 minutes 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.
ER130905_10.jpg Fig10: Flow cytometric analysis of alpha Tubulin was done on Hela cells. The cells were fixed, permeabilized and stained with the primary antibody (ER130905, 1/50) (blue). After incubation of the primary antibody at room temperature for an hour, the cells were stained with a Alexa Fluor®488 conjugate-Goat anti-Rabbit IgG Secondary antibody at 1/1000 dilution for 30 minutes.Unlabelled sample was used as a control (cells without incubation with primary antibody; red).
ER130905_11.jpg Fig11: Immunofluorescence analysis of paraffin-embedded human stomach tissue labeling alpha Tubulin with Rabbit anti-alpha Tubulin antibody (ER130905) at 1/200 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 (ER130905, green) at 1/200 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).
ER130905_12.jpg Fig12: Immunofluorescence analysis of paraffin-embedded mouse pancreas tissue labeling alpha Tubulin with Rabbit anti-alpha Tubulin antibody (ER130905) at 1/200 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 (ER130905, green) at 1/200 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).
ER130905_13.jpg Fig13: Immunofluorescence analysis of paraffin-embedded rat large intestine tissue labeling alpha Tubulin with Rabbit anti-alpha Tubulin antibody (ER130905) at 1/200 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 (ER130905, green) at 1/200 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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