Beta III Tubulin Mouse Monoclonal Antibody [A8-D10]
cat.: M0805-8
Product Type: Mouse monoclonal IgG2a, primary antibodies
Species reactivity: Human, Mouse, Rat
Applications: WB, IF-Cell, IHC-P, FC, IF-Tissue
Clonality: Monoclonal
Clone number: A8-D10
Form: Liquid
Storage condition: Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles.
Storage buffer: 1*PBS (pH7.4), 0.2% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
Concentration: 2ug/ul
Purification: Immunogen affinity purified.
Molecular weight: Predicted band size: 50 kDa
Isotype: IgG2a
Immunogen: Synthetic peptide (KLH-coupled) within human Tubulin beta-3 chain aa 401-450.
Positive control: SH-SY5Y cell lysate, U-87 MG cell lysate, A-172 cell lysate, Neuro-2a cell lysate, PC-12 cell lysate, mouse brain tissue lysate, rat brain tissue lysate, HEK-293, SH-SY5Y, Neuro-2a, human brain tissue, mouse brain tissue, rat brain tissue, mouse hippocampus tissue, MCF7.
Subcellular location: Cytoplasm. Cytoskeleton. Microtubule.
Recommended Dilutions:
  WB
  IF-Cell
  IHC-P
  FC
  IF-Tissue
1:2,000-1:5,000
1:500-1:1,000
1:2,000
1:1,000
1:200
Uniprot #: SwissProt: Q13509 Human | Q9ERD7 Mouse | Q4QRB4 Rat
Alternative names: beta 3 tubulin beta 4 beta-4 CDCBM CDCBM1 CFEOM3 CFEOM3A FEOM3 M(beta)3 M(beta)6 MC1R Neuron specific beta III Tubulin Neuron-specific class III beta-tubulin QccE-11995 QccE-15186 TBB3_HUMAN Tubb 3 TUBB3 TUBB4 Tubulin beta 3 Tubulin beta 3 chain Tubulin beta 4 Tubulin beta III Tubulin beta-3 chain Tubulin beta-4 chain Tubulin beta-III tuj 1 tuj1
Images
M0805-8_1.jpg Fig1: Western blot analysis of Beta III Tubulin on different lysates with Mouse anti-Beta III Tubulin antibody (M0805-8) at 1/2,000 dilution and competitor's antibody at 1/2,000 dilution.

Lane 1: SH-SY5Y cell lysate
Lane 2: U-87 MG cell lysate
Lane 3: A-172 cell lysate
Lane 4: Neuro-2a cell lysate
Lane 5: PC-12 cell lysate
Lane 6: Mouse brain tissue lysate
Lane 7: Rat brain tissue lysate

Lysates/proteins at 10 µg/Lane.

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

Exposure time: 11 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 (M0805-8) at 1/2,000 dilution and competitor's antibody at 1/2,000 dilution were used in 5% BSA at 4℃ overnight. Goat Anti-Mouse IgG - HRP Secondary Antibody (HA1006) at 1/50,000 dilution was used for 1 hour at room temperature.
M0805-8_2.jpg Fig2: Immunocytochemistry analysis of HEK-293 cells labeling Beta III Tubulin with Mouse anti-Beta III Tubulin antibody (M0805-8) 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 Mouse anti-Beta III Tubulin antibody (M0805-8) at 1/500 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Mouse IgG H&L (iFluor™ 488, HA1125) 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 (ET1602-4, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Rabbit IgG H&L (iFluor™ 594, HA1122) were used as the secondary antibody at 1/1,000 dilution.
M0805-8_3.jpg Fig3: Western blot analysis of Beta III Tubulin on different lysates with Mouse anti-Beta III Tubulin antibody (M0805-8) at 1/2,000 dilution.

Lane 1: SH-SY5Y cell lysate
Lane 2: U-87 MG cell lysate
Lane 3: A-172 cell lysate
Lane 4: Neuro-2a cell lysate
Lane 5: PC-12 cell lysate
Lane 6: Mouse brain tissue lysate
Lane 7: Rat brain tissue lysate

Lysates/proteins at 10 µg/Lane.

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

Exposure time: 11 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 (M0805-8) at 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Mouse IgG - HRP Secondary Antibody (HA1006) at 1/50,000 dilution was used for 1 hour at room temperature.
M0805-8_4.jpg Fig4: Immunocytochemistry analysis of SH-SY5Y cells labeling Beta III Tubulin with Mouse anti-Beta III Tubulin antibody (M0805-8) 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 Mouse anti-Beta III Tubulin antibody (M0805-8) at 1/500 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Mouse IgG H&L (iFluor™ 488, HA1125) 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 (ET1602-4, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Rabbit IgG H&L (iFluor™ 594, HA1122) were used as the secondary antibody at 1/1,000 dilution.
M0805-8_5.jpg Fig5: Immunocytochemistry analysis of Neuro-2a cells labeling Beta III Tubulin with Mouse anti-Beta III Tubulin antibody (M0805-8) 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 Mouse anti-Beta III Tubulin antibody (M0805-8) at 1/500 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Mouse IgG H&L (iFluor™ 488, HA1125) 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.
M0805-8_6.jpg Fig6: Immunohistochemical analysis of paraffin-embedded human brain tissue with Mouse anti-Beta III Tubulin antibody (M0805-8) at 1/2,000 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 (M0805-8) at 1/2,000 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.
M0805-8_7.jpg Fig7: Immunohistochemical analysis of paraffin-embedded mouse brain tissue with Mouse anti-Beta III Tubulin antibody (M0805-8) at 1/2,000 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 (M0805-8) at 1/2,000 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.
M0805-8_8.jpg Fig8: Immunohistochemical analysis of paraffin-embedded rat brain tissue with Mouse anti-Beta III Tubulin antibody (M0805-8) at 1/2,000 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 (M0805-8) at 1/2,000 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.
M0805-8_9.jpg Fig9: Immunofluorescence analysis of paraffin-embedded mouse brain tissue labeling Beta III Tubulin (M0805-8) and Synaptophysin (ET1606-56).

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 antibodies Beta III Tubulin (M0805-8, green) at 1/200 dilution and Synaptophysin (ET1606-56, red) at 1/200 dilution overnight at 4 ℃, washed with PBS.

Alexa Fluor® 488 conjugate-Goat anti-Mouse IgG and Alexa Fluor® 594 conjugate-Goat anti-Rabbit IgG were used as the secondary antibodies at 1/500 dilution. DAPI was used as nuclear counterstain.
M0805-8_10.jpg Fig10: Immunofluorescence analysis of paraffin-embedded mouse hippocampus tissue labeling Beta III Tubulin (M0805-8) and Synaptophysin (ET1606-56).

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 antibodies Beta III Tubulin (M0805-8, green) at 1/200 dilution and Synaptophysin (ET1606-56, red) at 1/200 dilution overnight at 4 ℃, washed with PBS.

Alexa Fluor® 488 conjugate-Goat anti-Mouse IgG and Alexa Fluor® 594 conjugate-Goat anti-Rabbit IgG were used as the secondary antibodies at 1/500 dilution. DAPI was used as nuclear counterstain.
M0805-8_11.jpg Fig11: Immunofluorescence analysis of paraffin-embedded rat brain tissue labeling Beta III Tubulin (M0805-8) and Synaptophysin (ET1606-56).

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 antibodies Beta III Tubulin (M0805-8, green) at 1/200 dilution and Synaptophysin (ET1606-56, red) at 1/200 dilution overnight at 4 ℃, washed with PBS.

Alexa Fluor® 488 conjugate-Goat anti-Mouse IgG and Alexa Fluor® 594 conjugate-Goat anti-Rabbit IgG were used as the secondary antibodies at 1/500 dilution. DAPI was used as nuclear counterstain.
M0805-8_12.jpg Fig12: Flow cytometric analysis of MCF7 cells labeling Beta III Tubulin.

Cells were fixed and permeabilized. Then stained with the primary antibody (M0805-8, 1/1,000) (red) compared with Mouse IgG1 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-Mouse IgG Secondary antibody (HA1125) 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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