WB
Western blot analysis of Beta III Tubulin on different lysates with Mouse anti-Beta III Tubulin 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. 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 at 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Mouse IgG - HRP Secondary Antibody at 1/50,000 dilution was used for 1 hour at room temperature.IHC
Immunohistochemical analysis of paraffin-embedded human brain tissue with Mouse anti-Beta III Tubulin antibody 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 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.ICC/IF
Immunocytochemistry analysis of SH-SY5Y cells labeling Beta III Tubulin with Mouse anti-Beta III Tubulin antibody 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 at 1/500 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Mouse IgG H&L (488) 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 (red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Rabbit IgG H&L (594) were used as the secondary antibody at 1/1,000 dilution.IF-P
Immunofluorescence analysis of paraffin-embedded mouse hippocampus tissue labeling Beta III Tubulin and Synaptophysin. 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 (green) at 1/200 dilution and Synaptophysin (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.FC
Flow cytometric analysis of MCF7 cells labeling Beta III Tubulin. Cells were fixed and permeabilized. Then stained with the primary antibody (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 at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).| Product Name | Beta III Tubulin Mouse Monoclonal Antibody |
|---|---|
| Antibody Type | Primary Antibodies |
| Immunogen | Synthetic peptide (KLH-coupled) within human Tubulin beta-3 chain aa 401-450. |
| Clonality | monoclonal |
|---|---|
| Isotype | IgG2a |
| Host Species | Mouse |
| Tested Applications | FCICC/IFIF-PIHCWB |
| WB:1:2000-1:5000 IHC:1:2000 ICC/IF:1:500-1:1000 IF-P: 1:200 FC:1:1000 |
|
| Species Reactivity | HumanMouseRat |
| Concentration | 1mg/ml |
| Purification | Affinity purified |
| Gene Symbol | TUBB3 |
|---|---|
| Gene Synonyms | CDCBM FEOM3 TUBB4 CDCBM1 CFEOM3 beta-4 CFEOM3A |
| Gene Full Name | tubulin beta 3 class III |
| Gene Summary | This gene encodes a class III member of the beta tubulin protein family. Beta tubulins are one of two core protein families (alpha and beta tubulins) that heterodimerize and assemble to form microtubules. This protein is primarily expressed in neurons and may be involved in neurogenesis and axon guidance and maintenance. Mutations in this gene are the cause of congenital fibrosis of the extraocular muscles type 3. Alternate splicing results in multiple transcript variants. A pseudogene of this gene is found on chromosome 6. [provided by RefSeq, Oct 2010] |
| Molecular Weight(MW) | 50kDa |
| Cellular Localization | Cytoplasm. Cytoskeleton. Microtubule. |
WB
Western blot analysis of Beta III Tubulin on different lysates with Mouse anti-Beta III Tubulin 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. 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 at 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Mouse IgG - HRP Secondary Antibody at 1/50,000 dilution was used for 1 hour at room temperature.
IHC
Immunohistochemical analysis of paraffin-embedded human brain tissue with Mouse anti-Beta III Tubulin antibody 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 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.
ICC/IF
Immunocytochemistry analysis of SH-SY5Y cells labeling Beta III Tubulin with Mouse anti-Beta III Tubulin antibody 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 at 1/500 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Mouse IgG H&L (488) 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 (red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Rabbit IgG H&L (594) were used as the secondary antibody at 1/1,000 dilution.
IF-P
Immunofluorescence analysis of paraffin-embedded mouse hippocampus tissue labeling Beta III Tubulin and Synaptophysin. 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 (green) at 1/200 dilution and Synaptophysin (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.
FC
Flow cytometric analysis of MCF7 cells labeling Beta III Tubulin. Cells were fixed and permeabilized. Then stained with the primary antibody (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 at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).| Application Notes | WB:1:2000-1:5000 IHC:1:2000 ICC/IF:1:500-1:1000 IF-P: 1:200 FC:1:1000 |
|---|
| Form | Liquid |
|---|---|
| Storage Instructions | 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. |
Data sheet for OM272431