Beta III Tubulin Mouse Monoclonal Antibody [A8-D10]
Catalog# M0805-8
Beta III Tubulin Mouse Monoclonal Antibody [A8-D10]
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WB
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IF-Cell
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IHC-P
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FC
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IF-Tissue
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Human
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Mouse
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Rat
Overview
Product Name
Beta III Tubulin Mouse Monoclonal Antibody [A8-D10]
Antibody Type
Mouse Monoclonal Antibody
Immunogen
Synthetic peptide (KLH-coupled) within human Tubulin beta-3 chain aa 401-450.
Species Reactivity
Human, Mouse, Rat
Validated Applications
WB, IF-Cell, IHC-P, FC, IF-Tissue
Molecular Weight
Predicted band size: 50 kDa
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.
Conjugation
unconjugated
Clone Number
A8-D10
RRID
Product Features
Form
Liquid
Concentration
2ug/ul
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.
Isotype
IgG2a
Purification Method
Immunogen affinity purified.
Application Dilution
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WB
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1:2,000-1:5,000
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IF-Cell
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1:500-1:1,000
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IHC-P
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1:2,000
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FC
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1:1,000
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IF-Tissue
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1:200
Applications in Publications
Species in Publications
Rat | See 3 publications below |
Mouse | See 1 publications below |
Human | See 1 publications below |
Target
Function
Tubulin is a compound of subunits of A tubulin and B tubulin. Class III beta tubulin (beta III-tubulin) is a vertebrate tubulin isotype specific to the neurons and mammalian testis cells, making it an ideal neuronal marker. Overexpression of class III beta tubulin is associated with the resistances of microtubule-targeted cancer drugs in lung cancer cell lines, breast cancer cell lines, and ovarian tumors.
Background References
1. Tischfield M A et al. Human TUBB3 mutations perturb microtubule dynamics, kinesin interactions, and axon guidance. Cell 140:74-87 (2010).
2. Fourest-Lieuvin A et al. Microtubule regulation in mitosis: tubulin phosphorylation by the cyclin-dependent kinase Cdk1. Mol Biol Cell 17:1041-1050 (2006).
Sequence Similarity
Belongs to the tubulin family.
Tissue Specificity
Expression is primarily restricted to central and peripheral nervous system. Greatly increased expression in most cancerous tissues.
Post-translational Modification
Some glutamate residues at the C-terminus are polyglutamylated, resulting in polyglutamate chains on the gamma-carboxyl group. Polyglutamylation plays a key role in microtubule severing by spastin (SPAST). SPAST preferentially recognizes and acts on microtubules decorated with short polyglutamate tails: severing activity by SPAST increases as the number of glutamates per tubulin rises from one to eight, but decreases beyond this glutamylation threshold.; Some glutamate residues at the C-terminus are monoglycylated but not polyglycylated due to the absence of functional TTLL10 in human. Monoglycylation is mainly limited to tubulin incorporated into axonemes (cilia and flagella). Both polyglutamylation and monoglycylation can coexist on the same protein on adjacent residues, and lowering glycylation levels increases polyglutamylation, and reciprocally. The precise function of monoglycylation is still unclear (Probable).; Phosphorylated on Ser-172 by CDK1 during the cell cycle, from metaphase to telophase, but not in interphase. This phosphorylation inhibits tubulin incorporation into microtubules.
Subcellular Location
Cytoplasm. Cytoskeleton. Microtubule.
Synonyms
beta 3 tubulin antibody
beta 4 antibody
beta-4 antibody
CDCBM antibody
CDCBM1 antibody
CFEOM3 antibody
CFEOM3A antibody
FEOM3 antibody
M(beta)3 antibody
M(beta)6 antibody
Expandbeta 3 tubulin antibody
beta 4 antibody
beta-4 antibody
CDCBM antibody
CDCBM1 antibody
CFEOM3 antibody
CFEOM3A antibody
FEOM3 antibody
M(beta)3 antibody
M(beta)6 antibody
MC1R antibody
Neuron specific beta III Tubulin antibody
Neuron-specific class III beta-tubulin antibody
QccE-11995 antibody
QccE-15186 antibody
TBB3_HUMAN antibody
Tubb 3 antibody
TUBB3 antibody
TUBB4 antibody
Tubulin beta 3 antibody
Tubulin beta 3 chain antibody
Tubulin beta 4 antibody
Tubulin beta III antibody
Tubulin beta-3 chain antibody
Tubulin beta-4 chain antibody
Tubulin beta-III antibody
tuj 1 antibody
tuj1 antibody
CollapseImages
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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. -
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. -
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. -
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. -
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. -
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. -
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. -
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. -
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. -
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. -
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. -
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).
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"
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