beta Tubulin Recombinant Rabbit Monoclonal Antibody [SR25-04] - BSA and Azide free
Catalog# HA750049
beta Tubulin Recombinant Rabbit Monoclonal Antibody [SR25-04] - BSA and Azide free
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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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Human
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Mouse
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Rat
Overview
Product Name
beta Tubulin Recombinant Rabbit Monoclonal Antibody [SR25-04] - BSA and Azide free
Antibody Type
Recombinant Rabbit monoclonal Antibody
Immunogen
Synthetic peptide within Human beta Tubulin aa 308-357 / 444.
Species Reactivity
Human, Mouse, Rat
Validated Applications
WB, IF-Cell, IHC-P, FC
Molecular Weight
Predicted band size: 50 kDa
Positive Control
HeLa cell lysate, 293T cell lysate, MCF7 cell lysate, SH-SY5Y cell lysate, U-2 OS cell lysate, Jurkat cell lysate, Neuro-2a cell lysate, NIH/3T3 cell lysate, PC-12 cell lysate, mouse brain tissue lysate, rat brain tissue lysate, SH-SY5Y, Hela, NIH/3T3, CRC, N2A, PC-12, human fallopian tube tissue, human colon carcinoma tissue, rat kidney tissue, mouse large intestine tissue.
Conjugation
unconjugated
Clone Number
SR25-04
Product Features
Form
Liquid
Concentration
1ug/ul
Storage Instructions
Store at +4℃ after thawing. Aliquot store at -20℃ or -80℃. Avoid repeated freeze / thaw cycles.
Storage Buffer
PBS (pH7.4).
Isotype
IgG
Purification Method
Protein A affinity purified.
Application Dilution
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WB
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1:20,000-1:100,000
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IF-Cell
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1:200-1:1,000
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IHC-P
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1:400-1:1,000
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FC
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1:500-1:1,000
Target
Function
Tubulin is one of several members of a small family of globular proteins. The most common members of the tubulin family are α-tubulin and β-tubulin. The beta-tubulin (relative molecular weight about 50 kDa) is counterpart of alpha-tubulin in tubulin heterodimer, it is coded by multiple tubulin genes and it is also posttranslationally modified. Heterogeneity of subunit is concentrated in C-terminal structural domain. Beta-Tubulin may have bound GTP or GDP. Under certain conditions β-tubulin can hydrolyze its bound GTP to GDP plus Pi, release the Pi, and exchange the GDP for GTP.
Background References
1. "Tumoral and tissue-specific expression of the major human beta-tubulin isotypes."Leandro-Garcia L.J., Leskela S., Landa I., Montero-Conde C., Lopez-Jimenez E., Leton R., .Cytoskeleton 67:214-223(2010).
2. "Five mouse tubulin isotypes and their regulated expression during development."Lewis S.A., Lee M.G.-S., Cowan N.J.J. Cell Biol. 101:852-861(1985).
Sequence Similarity
Belongs to the tubulin family.
Tissue Specificity
Ubiquitously expressed with highest levels in spleen, thymus and immature brain.
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
Synonyms
Beta 4 tubulin antibody
Beta 5 tubulin antibody
BetaTubulin antibody
TBB5_HUMAN antibody
TUBB antibody
TUBB2 antibody
TUBB2A antibody
TUBB5 antibody
tubulin beta 2A antibody
Tubulin beta chain antibody
ExpandImages
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Western blot analysis of beta Tubulin on different lysates with Rabbit anti-beta Tubulin antibody (HA750049) at 1/20,000 dilution.
Lane 1: HeLa cell lysate (15 µg/Lane)
Lane 2: 293T cell lysate (15 µg/Lane)
Lane 3: MCF7 cell lysate (15 µg/Lane)
Lane 4: SH-SY5Y cell lysate (15 µg/Lane)
Lane 5: U-2 OS cell lysate (15 µg/Lane)
Lane 6: Jurkat cell lysate (15 µg/Lane)
Lane 7: Neuro-2a cell lysate (15 µg/Lane)
Lane 8: NIH/3T3 cell lysate (15 µg/Lane)
Lane 9: PC-12 cell lysate (15 µg/Lane)
Lane 10: Mouse brain tissue lysate (20 µg/Lane)
Lane 11: Rat brain tissue lysate (20 µg/Lane)
Predicted band size: 50 kDa
Observed band size: 50 kDa
Exposure time: 1 minute 20 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 (HA750049) at 1/20,000 dilution was 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. -
Immunocytochemistry analysis of SH-SY5Y cells labeling beta Tubulin with Rabbit anti-beta Tubulin antibody (HA750049) at 1/200 dilution.
Cells were fixed in 4% paraformaldehyde for 10 minutes at 37 ℃, permeabilized with 0.05% Triton X-100 in PBS for 20 minutes, and then blocked with 2% negative goat serum for 30 minutes at room temperature. Cells were then incubated with Rabbit anti-beta Tubulin antibody (HA750049) at 1/200 dilution in 2% negative goat serum overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. Nuclear DNA was labelled in blue with DAPI.
Beta tubulin (M1305-2, red) was stained at 1/200 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution. -
Immunocytochemistry analysis of HeLa cells labeling beta Tubulin with Rabbit anti-beta Tubulin antibody (HA750049) at 1/500 dilution.
Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 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-beta Tubulin antibody (HA750049) 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 (HA601187, 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. -
Immunocytochemistry analysis of NIH/3T3 cells labeling beta Tubulin with Rabbit anti-beta Tubulin antibody (HA750049) at 1/500 dilution.
Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 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-beta Tubulin antibody (HA750049) 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 (HA601187, 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. -
Immunocytochemistry analysis of PC-12 cells labeling beta Tubulin with Rabbit anti-beta Tubulin antibody (HA750049) at 1/100 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-beta Tubulin antibody (HA750049) at 1/100 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. -
Immunohistochemical analysis of paraffin-embedded human colon carcinoma tissue with Rabbit anti-beta Tubulin antibody (HA750049) at 1/400 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 (HA750049) at 1/400 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 kidney tissue with Rabbit anti-beta Tubulin antibody (HA750049) at 1/400 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 (HA750049) at 1/400 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 large intestine tissue with Rabbit anti-beta Tubulin antibody (HA750049) at 1/400 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 (HA750049) at 1/400 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. -
Flow cytometric analysis of NIH/3T3 cells labeling beta Tubulin.
Cells were fixed and permeabilized. Then stained with the primary antibody (HA750049, 1ug/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).
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"
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