Beta Tubulin Monoclonal antibody

Beta Tubulin Monoclonal Antibody for WB, IHC, IF/ICC, IF-P, IP, ELISA

Host / Isotype

Mouse / IgG2a

Reactivity

human, mouse, rat, pig, zebrafish, nematode and More (5)

Applications

WB, IHC, IF/ICC, IF-P, IP, ELISA

Conjugate

Unconjugated

CloneNo.

1D4A4

Cat no : 66240-1-Ig

Synonyms

Tubulin beta III, Tubulin beta 3 chain, Tubulin beta-4 chain, Tubulin beta-3 chain, TUBB4


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  • Western Blot (WB) analysis of various lysates using Beta Tubulin Monoclonal antibody (66240-1-Ig)

    WB analysis using 66240-1-Ig

    Various lysates were subjected to SDS PAGE followed by western blot with 66240-1-Ig (beta Tubulin antibody) at dilution of 1:200000 incubated at room temperature for 1.5 hours.

  • Western Blot (WB) analysis of various lysates using Beta Tubulin Monoclonal antibody (66240-1-Ig)

    WB analysis using 66240-1-Ig

    Various lysates were subjected to SDS PAGE followed by western blot with 66240-1-Ig (beta Tubulin antibody) at dilution of 1:100000 incubated at room temperature for 1.5 hours.

  • Immunoprecipitation (IP) experiment of HeLa cells using Beta Tubulin Monoclonal antibody (66240-1-Ig)

    IP experiment of HeLa using 66240-1-Ig

    IP result of anti-Beta Tubulin (IP:66240-1-Ig, 5ug; Detection:66240-1-Ig 1:1000) with HeLa cells lysate 880ug.

  • Immunohistochemistry (IHC) staining of human brain tissue using Beta Tubulin Monoclonal antibody (66240-1-Ig)

    IHC staining of human brain using 66240-1-Ig

    Immunohistochemical analysis of paraffin-embedded human brain tissue slide using 66240-1-Ig (beta Tubulin antibody at dilution of 1:400 (under 10x lens).

  • Immunohistochemistry (IHC) staining of human brain tissue using Beta Tubulin Monoclonal antibody (66240-1-Ig)

    IHC staining of human brain using 66240-1-Ig

    Immunohistochemical analysis of paraffin-embedded human brain tissue slide using 66240-1-Ig (beta Tubulin antibody at dilution of 1:400 (under 40x lens).

  • Immunohistochemistry (IHC) staining of human breast cancer tissue using Beta Tubulin Monoclonal antibody (66240-1-Ig)

    IHC staining of human breast cancer using 66240-1-Ig

    Immunohistochemical analysis of paraffin-embedded human breast cancer tissue slide using 66240-1-Ig (beta Tubulin antibody at dilution of 1:400 (under 10x lens).

  • Immunohistochemistry (IHC) staining of human breast cancer tissue using Beta Tubulin Monoclonal antibody (66240-1-Ig)

    IHC staining of human breast cancer using 66240-1-Ig

    Immunohistochemical analysis of paraffin-embedded human breast cancer tissue slide using 66240-1-Ig (beta Tubulin antibody at dilution of 1:400 (under 40x lens).

  • Immunohistochemistry (IHC) staining of human breast cancer tissue using Beta Tubulin Monoclonal antibody (66240-1-Ig)

    IHC staining of human breast cancer using 66240-1-Ig

    Immunohistochemical analysis of paraffin-embedded human breast cancer tissue slide using 66240-1-Ig (beta Tubulin antibody) at dilution of 1:10000 (under 10x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

  • Immunohistochemistry (IHC) staining of rat brain tissue using Beta Tubulin Monoclonal antibody (66240-1-Ig)

    IHC staining of rat brain using 66240-1-Ig

    Immunohistochemical analysis of paraffin-embedded rat brain tissue slide using 66240-1-Ig (beta Tubulin antibody at dilution of 1:400 (under 10x lens).

  • Immunohistochemistry (IHC) staining of rat brain tissue using Beta Tubulin Monoclonal antibody (66240-1-Ig)

    IHC staining of rat brain using 66240-1-Ig

    Immunohistochemical analysis of paraffin-embedded rat brain tissue slide using 66240-1-Ig (beta Tubulin antibody at dilution of 1:400 (under 40x lens).

  • Immunofluorescence (IF) / fluorescent staining of mouse eye tissue using Beta Tubulin Monoclonal antibody (66240-1-Ig)

    IF Staining of mouse eye using 66240-1-Ig

    Immunofluorescent analysis of (4% PFA) fixed paraffin-embedded mouse eye tissue using Beta Tubulin antibody (66240-1-Ig, Clone: 1D4A4 ) at dilution of 1:800 and CoraLite®488-Conjugated Goat Anti-Mouse IgG(H+L) (SA00013-1). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

  • Immunofluorescence (IF) / fluorescent staining of HeLa cells using Beta Tubulin Monoclonal antibody (66240-1-Ig)

    IF Staining of HeLa using 66240-1-Ig

    Immunofluorescent analysis of (4% PFA) fixed HeLa cells using 66240-1-Ig (beta Tubulin antibody), at dilution of 1:400 and CoraLite®488-Conjugated AffiniPure Goat Anti-Mouse IgG(H+L). Red: staning with CoraLite555-Phalloidin.

  • Immunofluorescence (IF) / fluorescent staining of HeLa cells using Beta Tubulin Monoclonal antibody (66240-1-Ig)

    IF Staining of HeLa using 66240-1-Ig

    Immunofluorescent analysis of (4% PFA) fixed HeLa cells using Beta Tubulin antibody (66240-1-Ig, Clone: 1D4A4 ) at dilution of 1:400 and CoraLite®488-Conjugated AffiniPure Goat Anti-Mouse IgG(H+L), CL594-Phalloidin (red).

  • Immunofluorescence (IF) / fluorescent staining of HepG2 cells using Beta Tubulin Monoclonal antibody (66240-1-Ig)

    IF Staining of HepG2 using 66240-1-Ig

    Immunofluorescent analysis of HepG2 cells using 66240-1-Ig (beta Tubulin antibody) at dilution of 1:50 and Alexa Fluor 488-conjugated AffiniPure Goat Anti-Mouse IgG (H+L).

  • Immunofluorescence (IF) / fluorescent staining of HepG2 cells using Beta Tubulin Monoclonal antibody (66240-1-Ig)

    IF Staining of HepG2 using 66240-1-Ig

    Immunofluorescent analysis of HepG2 cells using 66240-1-Ig (beta Tubulin antibody) at dilution of 1:50 and Alexa Fluor 488-conjugated AffiniPure Goat Anti-Mouse IgG (H+L).

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Tested Applications

Positive WB detected inHeLa cells, HEK-293 cells, HepG2 cells, Jurkat cells, HSC-T6 cells, MCF-7 cells, NCCIT cells, HT-1080 cells, LNCaP cells, ROS1728 cells, NIH/3T3 cells
Positive IP detected inHeLa cells
Positive IHC detected inhuman brain tissue, human breast cancer tissue, rat brain tissue
Note: suggested antigen retrieval with TE buffer pH 9.0; (*) Alternatively, antigen retrieval may be performed with citrate buffer pH 6.0
Positive IF-P detected inmouse eye tissue
Positive IF/ICC detected inHeLa cells, HepG2 cells

Recommended dilution

ApplicationDilution
Western Blot (WB)WB : 1:20000-1:100000
Immunoprecipitation (IP)IP : 0.5-4.0 ug for 1.0-3.0 mg of total protein lysate
Immunohistochemistry (IHC)IHC : 1:20-1:1000
Immunofluorescence (IF)-PIF-P : 1:400-1:1600
Immunofluorescence (IF)/ICCIF/ICC : 1:200-1:800
It is recommended that this reagent should be titrated in each testing system to obtain optimal results.
Sample-dependent, Check data in validation data gallery.

Product Information

66240-1-Ig targets Beta Tubulin in WB, IHC, IF/ICC, IF-P, IP, ELISA applications and shows reactivity with human, mouse, rat, pig, zebrafish, nematode samples.

Tested Reactivity human, mouse, rat, pig, zebrafish, nematode
Cited Reactivityhuman, mouse, rat, pig, rabbit, canine, monkey, chicken, zebrafish, sheep
Host / Isotype Mouse / IgG2a
Class Monoclonal
Type Antibody
Immunogen Beta Tubulin fusion protein Ag0117 相同性解析による交差性が予測される生物種
Full Name tubulin, beta 3
Calculated molecular weight 450 aa, 50 kDa
Observed molecular weight 50-55 kDa
GenBank accession numberBC000748
Gene symbol TUBB3
Gene ID (NCBI) 10381
RRIDAB_2881629
Conjugate Unconjugated
Form Liquid
Purification MethodProtein A purification
Storage Buffer PBS with 0.02% sodium azide and 50% glycerol pH 7.3.
Storage ConditionsStore at -20°C. Stable for one year after shipment. Aliquoting is unnecessary for -20oC storage.

Background Information

There are five tubulins in human cells: alpha, beta, gamma, delta, and epsilon. Tubulins are conserved across species. They form heterodimers, which multimerize to form a microtubule filament. An alpha and beta tubulin heterodimer is the basic structural unit of microtubules. The alpha and beta tubulins (+/- 55 kDa MW) are homologous but are not identical. Beta tubulins have been widely used as loading control.

 

What is the molecular weight of beta-tubulin? Are there any isoforms of beta-tubulin?

The molecular weight of tubulin is 50-52 kDa. Humans have eight beta-tubulin isotypes, encoded by different genes, that differ in their C-terminal sequences. They have different tissue expression profiles and can rise to microtubules of different properties (PMID: 20191564).

 

How to use beta-tubulin as a loading control

Beta-tubulin is one of the most commonly used references as a loading control for cell lysates in western blotting. It is abundantly expressed across various tissues and developmental stages and highly conserved across species. However, since some variability has been observed in the expression levels of commonly used housekeeping genes (PMID: 15627964), it is recommended that more than one loading control antibody is used while developing new assays. More information can be found here: https://www.ptglab.com/news/blog/loading-control-antibodies-for-western-blotting/.

 

What drugs can influence beta-tubulin and organization of microtubules?

Many drugs that affect microtubule dynamics target beta-tubulin, mainly by interfering with the GTP hydrolysis (PMID: 21381049). Paclitaxel (Taxol) is used to stabilize microtubules by slowing down their depolymerization, while colchicine and vinca alkaloids (vinblastine) destabilize microtubules. They are used in research and also in the clinic as anti-cancer agents.

 

Is beta-tubulin post-translationally modified?

Yes, tubulins are subject to extensive post-translational modifications (PTMs) that affect the organization of microtubules and their dynamics. The most common modifications include polyglutamylation, polyglycylation, polyamination, glycososylation, glycation, phosphorylation, and acetylation (PMID: 24801181 and 25468068).

Protocols

Product Specific Protocols
WB protocol for Beta Tubulin antibody 66240-1-IgDownload protocol
IHC protocol for Beta Tubulin antibody 66240-1-IgDownload protocol
IF protocol for Beta Tubulin antibody 66240-1-IgDownload protocol
IP protocol for Beta Tubulin antibody 66240-1-IgDownload protocol
Standard Protocols
Click here to view our Standard Protocols

Publications

SpeciesApplicationTitle
humanIF

Nat Biotechnol

Magnify is a universal molecular anchoring strategy for expansion microscopy

Authors - Aleksandra Klimas
humanIF

Nature

ER proteins decipher the tubulin code to regulate organelle distribution

Authors - Pengli Zheng
WB

Nature

IL-27 signalling promotes adipocyte thermogenesis and energy expenditure.

Authors - Qian Wang
humanWB

Science

A prometastatic splicing program regulated by SNRPA1 interactions with structured RNA elements.

Authors - Lisa Fish
WB

Adv Mater

A Physiologically Responsive Nanocomposite Hydrogel for Treatment of Head and Neck Squamous Cell Carcinoma via Proteolysis-targeting Chimeras Enhanced Immunotherapy

Authors - Yaping Wu
humanWB

Gut

Integrative multiomics enhancer activity profiling identifies therapeutic vulnerabilities in cholangiocarcinoma of different etiologies

Authors - Jing Han Hong

Reviews

The reviews below have been submitted by verified Proteintech customers who received an incentive forproviding their feedback.


FH

Aditya (Verified Customer) (11-01-2024)

great with flourescence, our standard housekeeping control

  • Applications: Western Blot
FH

P (Verified Customer) (09-23-2024)

Excellent

  • Applications: Western Blot
  • Primary Antibody Dilution: 2000
  • Cell Tissue Type: HEK
Beta Tubulin Antibody Western Blot validation (2000 dilution) in HEK (Cat no:66240-1-Ig)
FH

Moritz (Verified Customer) (09-12-2024)

The antibody worked very well and gave a very strong signal (can be diluted more). Used it in HEK293T cell lysate analysed with WB.

  • Applications: Western Blot
  • Primary Antibody Dilution: 1:20000
  • Cell Tissue Type: HEK293T
Beta Tubulin Antibody Western Blot validation (1:20000 dilution) in HEK293T (Cat no:66240-1-Ig)
FH

Tsimafei (Verified Customer) (08-04-2024)

Excellent antibody! The membrane was incubated for 1h at RT

  • Applications: Western Blot
  • Primary Antibody Dilution: 1:20000
  • Cell Tissue Type: mESCs
Beta Tubulin Antibody Western Blot validation (1:20000 dilution) in mESCs (Cat no:66240-1-Ig)
FH

Andrea (Verified Customer) (01-11-2023)

Antibody were incubated for 2h at room temperature. Provide a specific tubulin signal

  • Applications: Western Blot
  • Primary Antibody Dilution: 1:10000
  • Cell Tissue Type: mESCs
Beta Tubulin Antibody Western Blot validation (1:10000 dilution) in mESCs (Cat no:66240-1-Ig)
FH

Emma (Verified Customer) (11-29-2021)

Very clean antibody and runs at the right size by western blot.

  • Applications: Western Blot
  • Primary Antibody Dilution: 1:10000
  • Cell Tissue Type: Prostate cell lines (LNCaP, CWR22Rv1, DU145, PC3)
FH

Raul (Verified Customer) (08-20-2021)

Very good antibody producing a single strong band at ~55kDa

  • Applications: Western Blot
  • Primary Antibody Dilution: 1:20000
  • Cell Tissue Type: SW1353 chondrosarcorma cell line
Beta Tubulin Antibody Western Blot validation (1:20000 dilution) in SW1353 chondrosarcorma cell line (Cat no:66240-1-Ig)
FH

Yu Siong (Verified Customer) (08-18-2020)

Western blot of 4 bands: 4 different treatments.

  • Applications: Western Blot, Cell culture
  • Primary Antibody Dilution: 1:5000
  • Cell Tissue Type: primary porcine brain endothelial cells
Beta Tubulin Antibody Western Blot,Cell culture validation (1:5000 dilution) in primary porcine brain endothelial cells (Cat no:66240-1-Ig)
FH

Kathryn (Verified Customer) (12-10-2019)

Used 1:20000 in 5% BSA on a small amount of neonatal mouse ovary. Very clean and specific, and great that it works at such a low concentration.

  • Applications: Western Blot,
  • Primary Antibody Dilution: 1:20000
  • Cell Tissue Type: mouse ovary
Beta Tubulin Antibody Western Blot, validation (1:20000 dilution) in mouse ovary (Cat no:66240-1-Ig)
FH

Sara (Verified Customer) (10-10-2019)

This antibody works great in WB. We use it as loading control in whole cell lysates (mice B cell line), 1:10.000 O/N at 4°C.

  • Applications: Western Blot,
  • Primary Antibody Dilution: 1:10.000
  • Cell Tissue Type: mice, B lymphocytes
FH

VLADIMIR (Verified Customer) (09-25-2019)

B-tubulin IgG2a were used to prepare Fab' conjugated with Alexa 647-maleimide

  • Applications: Immunofluorescence,
  • Cell Tissue Type: HeLa
FH

Jeffery (Verified Customer) (08-01-2019)

Very clean antibody. Works exceptionally well. Runs right at the 50kd marker.

  • Applications: Western Blot,
  • Primary Antibody Dilution: 1:20,000
  • Cell Tissue Type: Ins-1 rat beta cells
Beta Tubulin Antibody Western Blot, validation (1:20,000 dilution) in Ins-1 rat beta cells (Cat no:66240-1-Ig)
FH

Shashirekha (Verified Customer) (05-09-2019)

Works great!

  • Applications: Western Blot,
  • Primary Antibody Dilution: 1:5000
  • Cell Tissue Type: NSC-34
Beta Tubulin Antibody Western Blot, validation (1:5000 dilution) in NSC-34 (Cat no:66240-1-Ig)
FH

Zunair (Verified Customer) (12-03-2018)

  • Applications: Western Blot,
  • Primary Antibody Dilution: 1:5000
  • Cell Tissue Type: 786-O [786-0] (ATCC® CRL-1932™)
Beta Tubulin Antibody Western Blot, validation (1:5000 dilution) in 786-O [786-0] (ATCC® CRL-1932™) (Cat no:66240-1-Ig)