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beta Tubulin Mouse Monoclonal Antibody [A1-A4]

Specification

Quota

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BIO-STATION LIMITED

Contact: Sanry CHEN

Email: info@bio-station.net

Phone: 852-35650223

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Catalog# M1305-2

beta Tubulin Mouse Monoclonal Antibody [A1-A4]

Application

  • WB

  • IF-Cell

  • IHC-P

  • FC

Reactivity

  • Human

  • Mouse

  • Rat

This product has been cited in peer reviewed publications, see list HERE

Overview

Product Name

beta Tubulin Mouse Monoclonal Antibody [A1-A4]

Antibody Type

Mouse Monoclonal Antibody

Immunogen

Synthetic peptide within Human Beta tubulin aa 151-200 / 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 lysates, NIH/3T3 cell lysates, PC-12 cell lysates, SKOV-3 cells, HeLa, 293, human tonsil tissue, human spleen tissue, human colon tissue, rat brain tissue, NIH/3T3.

Conjugation

unconjugated

Clone Number

A1-A4

RRID

AB_3073058

Product Features

Form

Liquid

Concentration

2ug/ul

Storage Instructions

Shipped at 4℃. Store at +4℃ short term (1-2 weeks). It is recommended to aliquot into single-use upon delivery. Store at -20℃ long term.

Storage Buffer

1*PBS (pH7.4), 0.2% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.

Isotype

IgG3

Purification Method

Protein A affinity purified.

Application Dilution

  • WB

  • 1:5,000-1:20,000

  • IF-Cell

  • 1:100-1:500

  • IHC-P

  • 1:50-1:200

  • FC

  • 1:100-1:1,000

Target

Function

Tubulins is one of several members of a small family of globular proteins. The most common members of the tubulins family are α-tubulins and β-tubulins. The beta-tubulins (relative molecular weight about 50 kDa) is counterpart of alpha-tubulin in tubulins heterodimer, it is coded by multiple tubulins genes and it is also posttranslationally modified. Heterogeneity of subunit is concentrated in C-terminal structural domain. Beta-Tubulins 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, Cytoskeleton, Microtubule.

UNIPROT

SwissProt: P07437 Human

SwissProt: P99024 Mouse

SwissProt: P69897 Rat

Synonyms

beta 3 tubulin antibody

beta-4 antibody

CDCBM antibody

CDCBM1 antibody

CFEOM3 antibody

CFEOM3A antibody

FEOM3 antibody

M(beta)3 antibody

M(beta)6 antibody

MC1R antibody

Expand

Images

  • Western blot analysis of beta Tubulin on Hela cell lysates with Mouse anti-beta Tubulin antibody (<a href="/products/M1305-2" style="font-weight: bold;text-decoration: underline;">M1305-2</a>).<br /><br />Hela cell lysates at 10 µg/Lane.<br /><br />Predicted band size: 50 kDa<br />Observed band size: 50 kDa<br /><br />8% SDS-PAGE gel.<br /><br />Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (<a href="/products/M1305-2" style="font-weight: bold;text-decoration: underline;">M1305-2</a>) at serial dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Mouse IgG - HRP Secondary Antibody (<a href="/products/HA1006" style="font-weight: bold;text-decoration: underline;">HA1006</a>) at 1:100,000 dilution was used for 1 hour at room temperature.

    Western blot analysis of beta Tubulin on Hela cell lysates with Mouse anti-beta Tubulin antibody (M1305-2).

    Hela cell lysates at 10 µg/Lane.

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

    8% 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 (M1305-2) at serial dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Mouse IgG - HRP Secondary Antibody (HA1006) at 1:100,000 dilution was used for 1 hour at room temperature.

  • Western blot analysis of beta Tubulin on NIH/3T3 cell lysates with Mouse anti-beta Tubulin antibody (<a href="/products/M1305-2" style="font-weight: bold;text-decoration: underline;">M1305-2</a>).<br /><br />NIH/3T3 cell lysates at 10 µg/Lane.<br /><br />Predicted band size: 50 kDa<br />Observed band size: 50 kDa<br /><br />8% SDS-PAGE gel.<br /><br />Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (<a href="/products/M1305-2" style="font-weight: bold;text-decoration: underline;">M1305-2</a>) at serial dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Mouse IgG - HRP Secondary Antibody (<a href="/products/HA1006" style="font-weight: bold;text-decoration: underline;">HA1006</a>) at 1:100,000 dilution was used for 1 hour at room temperature.

    Western blot analysis of beta Tubulin on NIH/3T3 cell lysates with Mouse anti-beta Tubulin antibody (M1305-2).

    NIH/3T3 cell lysates at 10 µg/Lane.

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

    8% 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 (M1305-2) at serial dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Mouse IgG - HRP Secondary Antibody (HA1006) at 1:100,000 dilution was used for 1 hour at room temperature.

  • Western blot analysis of beta Tubulin on PC-12 cell lysates with Mouse anti-beta Tubulin antibody (<a href="/products/M1305-2" style="font-weight: bold;text-decoration: underline;">M1305-2</a>).<br /><br />PC-12 cell lysates at 10 µg/Lane.<br /><br />Predicted band size: 50 kDa<br />Observed band size: 50 kDa<br /><br />8% SDS-PAGE gel.<br /><br />Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (<a href="/products/M1305-2" style="font-weight: bold;text-decoration: underline;">M1305-2</a>) at serial dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Mouse IgG - HRP Secondary Antibody (<a href="/products/HA1006" style="font-weight: bold;text-decoration: underline;">HA1006</a>) at 1:100,000 dilution was used for 1 hour at room temperature.

    Western blot analysis of beta Tubulin on PC-12 cell lysates with Mouse anti-beta Tubulin antibody (M1305-2).

    PC-12 cell lysates at 10 µg/Lane.

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

    8% 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 (M1305-2) at serial dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Mouse IgG - HRP Secondary Antibody (HA1006) at 1:100,000 dilution was used for 1 hour at room temperature.

  • Western blot analysis of beta Tubulin on different lysates with Mouse anti-beta Tubulin antibody (<a href="/products/M1305-2" style="font-weight: bold;text-decoration: underline;">M1305-2</a>) at 1/1,000 dilution.<br /><br />Lane 1: HeLa cell lysate<br />Lane 2: HepG2 cell lysate<br />Lane 3: NIH/3T3 cell lysate<br />Lane 4: PC-12 cell lysate<br />Lane 5: C6 cell lysate<br />Lane 6: Vero cell lysate<br />Lane 7: COS-1 cell lysate<br /><br />Lysates/proteins at 20 µg/Lane.<br /><br />Predicted band size: 52 kDa<br />Observed band size: 52 kDa<br /><br />Exposure time: 2 seconds;<br /><br />4-20% SDS-PAGE gel.<br /><br />Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (<a href="/products/M1305-2" style="font-weight: bold;text-decoration: underline;">M1305-2</a>) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Mouse IgG - HRP Secondary Antibody (<a href="/products/HA1006" style="font-weight: bold;text-decoration: underline;">HA1006</a>) at 1/50,000 dilution was used for 1 hour at room temperature.

    Western blot analysis of beta Tubulin on different lysates with Mouse anti-beta Tubulin antibody (M1305-2) at 1/1,000 dilution.

    Lane 1: HeLa cell lysate
    Lane 2: HepG2 cell lysate
    Lane 3: NIH/3T3 cell lysate
    Lane 4: PC-12 cell lysate
    Lane 5: C6 cell lysate
    Lane 6: Vero cell lysate
    Lane 7: COS-1 cell lysate

    Lysates/proteins at 20 µg/Lane.

    Predicted band size: 52 kDa
    Observed band size: 52 kDa

    Exposure time: 2 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 (M1305-2) at 1/1,000 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 HeLa cells labeling beta Tubulin with Mouse anti-beta Tubulin antibody (<a href="/products/M1305-2" style="font-weight: bold;text-decoration: underline;">M1305-2</a>) at 1/100 dilution.<br /><br />Cells were fixed in 100% precooled methanol 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 Tubulin antibody (<a href="/products/M1305-2" style="font-weight: bold;text-decoration: underline;">M1305-2</a>) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Mouse IgG H&L (iFluor&trade; 594, <a href="/products/HA1126" style="font-weight: bold;text-decoration: underline;">HA1126</a>) 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.

    Immunocytochemistry analysis of HeLa cells labeling beta Tubulin with Mouse anti-beta Tubulin antibody (M1305-2) at 1/100 dilution.

    Cells were fixed in 100% precooled methanol 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 Tubulin antibody (M1305-2) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) 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.

  • Immunocytochemistry analysis of 293 cells labeling beta Tubulin with Mouse anti-beta Tubulin antibody (<a href="/products/M1305-2" style="font-weight: bold;text-decoration: underline;">M1305-2</a>) at 1/100 dilution.<br /><br />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 Tubulin antibody (<a href="/products/M1305-2" style="font-weight: bold;text-decoration: underline;">M1305-2</a>) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Mouse IgG H&L (iFluor&trade; 594, <a href="/products/HA1126" style="font-weight: bold;text-decoration: underline;">HA1126</a>) 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.

    Immunocytochemistry analysis of 293 cells labeling beta Tubulin with Mouse anti-beta Tubulin antibody (M1305-2) 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 Mouse anti-beta Tubulin antibody (M1305-2) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) 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 tonsil tissue using anti-beta tubulin antibody. Counter stained with hematoxylin.

    Immunohistochemical analysis of paraffin-embedded human tonsil tissue using anti-beta tubulin antibody. Counter stained with hematoxylin.

  • Immunohistochemical analysis of paraffin-embedded human spleen tissue using anti-beta tubulin antibody. Counter stained with hematoxylin.

    Immunohistochemical analysis of paraffin-embedded human spleen tissue using anti-beta tubulin antibody. Counter stained with hematoxylin.

  • Immunohistochemical analysis of paraffin-embedded human colon tissue using anti-beta tubulin antibody. Counter stained with hematoxylin.

    Immunohistochemical analysis of paraffin-embedded human colon tissue using anti-beta tubulin antibody. Counter stained with hematoxylin.

  • Immunohistochemical analysis of paraffin-embedded rat brain tissue using anti-beta tubulin antibody. Counter stained with hematoxylin.

    Immunohistochemical analysis of paraffin-embedded rat brain tissue using anti-beta tubulin antibody. Counter stained with hematoxylin.

  • Flow cytometric analysis of HeLa cells labeling beta Tubulin.<br /><br />Cells were fixed and permeabilized. Then stained with the primary antibody (<a href="/products/M1305-2" style="font-weight: bold;text-decoration: underline;">M1305-2</a>, 1μg/mL) (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&trade; 488 conjugate-Goat anti-Mouse IgG Secondary antibody (<a href="/products/HA1125" style="font-weight: bold;text-decoration: underline;">HA1125</a>) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

    Flow cytometric analysis of HeLa cells labeling beta Tubulin.

    Cells were fixed and permeabilized. Then stained with the primary antibody (M1305-2, 1μg/mL) (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).

  • Flow cytometric analysis of NIH/3T3 cells labeling beta Tubulin.<br /><br />Cells were fixed and permeabilized. Then stained with the primary antibody (<a href="/products/M1305-2" style="font-weight: bold;text-decoration: underline;">M1305-2</a>, 1μg/mL) (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&trade; 488 conjugate-Goat anti-Mouse IgG Secondary antibody (<a href="/products/HA1125" style="font-weight: bold;text-decoration: underline;">HA1125</a>) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

    Flow cytometric analysis of NIH/3T3 cells labeling beta Tubulin.

    Cells were fixed and permeabilized. Then stained with the primary antibody (M1305-2, 1μg/mL) (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).

  • Immunocytochemistry analysis of PC-12 cells labeling beta Tubulin with Mouse anti-beta Tubulin antibody (<a href="/products/M1305-2" style="font-weight: bold;text-decoration: underline;">M1305-2</a>) at 1/100 dilution.<br /><br />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 Tubulin antibody (<a href="/products/M1305-2" style="font-weight: bold;text-decoration: underline;">M1305-2</a>) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Mouse IgG H&L (iFluor&trade; 488, <a href="/products/HA1125" style="font-weight: bold;text-decoration: underline;">HA1125</a>) 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.<br /><br />beta Tubulin (<a href="/products/ET1602-4" style="font-weight: bold;text-decoration: underline;">ET1602-4</a>, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Rabbit IgG H&L (iFluor&trade; 594, <a href="/products/HA1122" style="font-weight: bold;text-decoration: underline;">HA1122</a>) were used as the secondary antibody at 1/1,000 dilution.

    Immunocytochemistry analysis of PC-12 cells labeling beta Tubulin with Mouse anti-beta Tubulin antibody (M1305-2) 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 Mouse anti-beta Tubulin antibody (M1305-2) at 1/100 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.

  • Flow cytometric analysis of PC-12 HeLa cells labeling beta Tubulin.<br /><br />Cells were fixed and permeabilized. Then stained with the primary antibody (<a href="/products/M1305-2" style="font-weight: bold;text-decoration: underline;">M1305-2</a>, 1μg/mL) (red) compared with Mouse IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor&trade; 488 conjugate-Goat anti-Mouse IgG Secondary antibody (<a href="/products/HA1125" style="font-weight: bold;text-decoration: underline;">HA1125</a>) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

    Flow cytometric analysis of PC-12 HeLa cells labeling beta Tubulin.

    Cells were fixed and permeabilized. Then stained with the primary antibody (M1305-2, 1μg/mL) (red) compared with Mouse 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-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"

Citation

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  1. WB
  2. COIP
  3. IHC

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