CoraLite® Plus 488-conjugated Beta Tubulin Monoclonal antibody

Beta Tubulin Monoclonal Antibody for WB, IF/ICC, FC (Intra)

Host / Isotype

Mouse / IgG2a

Reactivity

human, mouse, rat, pig, zebrafish, nematode

Applications

WB, IF/ICC, FC (Intra)

Conjugate

CoraLite® Plus 488 Fluorescent Dye

Publications

15

CloneNo.

1D4A4

Cat no : CL488-66240

Synonyms

TUBB3, Tubulin beta III, Tubulin beta 3 chain, Tubulin beta-3 chain, TUBB4

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Validation Data Gallery

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  WB analysis using CL488-66240

  Various lysates were subjected to SDS PAGE followed by western blot with CL488-66240 (Beta Tubulin antibody) at dilution of 1:5000 incubated at room temperature for 1.5 hours.

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  IF Staining of HepG2 using CL488-66240

  Immunofluorescent analysis of (4% PFA) fixed HepG2 cells using CL488-66240 (beta Tubulin antibody) at dilution of 1:100.

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  FC experiment of HeLa using CL488-66240

  1X10^6 HeLa cells were intracellularly stained with 0.4 ug CoraLite® Plus 488 Anti-Human Beta Tubulin (CL488-66240, Clone:1D4A4) (red), or 0.4 ug Mouse IgG2a Isotype Control (CL488-66360-2, Clone: K11A1B2A2) (blue). Cells were fixed with 4% PFA and permeabilized with Flow Cytometry Perm Buffer (PF00011-C).

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Tested Applications

• Positive WB detected in: HeLa cells, HEK-293 cells, HSC-T6 cells, NIH/3T3 cells
• Positive IF/ICC detected in: HepG2 cells
• Positive FC (Intra) detected in: HeLa cells

Recommended dilution

• Western Blot (WB): WB : 1:2000-1:10000
• Immunofluorescence (IF)/ICC: IF/ICC : 1:50-1:500
• Flow Cytometry (FC) (INTRA): FC (INTRA) : 0.40 ug per 10^6 cells in a 100 µl suspension

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.

Published Applications

• WB: See 2 publications below
• IF: See 12 publications below

Product Information

CL488-66240 targets Beta Tubulin in WB, IF/ICC, FC (Intra) applications and shows reactivity with human, mouse, rat, pig, zebrafish, nematode samples.

• Tested Reactivity: human, mouse, rat, pig, zebrafish, nematode
• Cited Reactivity: human, mouse, rat
• Host / Isotype: Mouse / IgG2a
• Class: Monoclonal
• Type: Antibody
• Immunogen: Beta Tubulin fusion protein Ag0117
• Full Name: tubulin, beta 3
• Calculated Molecular Weight: 450 aa, 50 kDa
• GenBank Accession Number: BC000748
• Gene Symbol: TUBB3
• Gene ID (NCBI): 10381
• RRID: AB_2883292
• Conjugate: CoraLite® Plus 488 Fluorescent Dye
• Excitation/Emission Maxima Wavelengths: 493 nm / 522 nm
• Form: Liquid
• Purification Method: Protein A purification
• Storage Buffer: PBS with 50% Glycerol, 0.05% Proclin300, 0.5% BSA, pH 7.3.
• Storage Conditions: Store at -20°C. Avoid exposure to light. Stable for one year after shipment. Aliquoting is unnecessary for -20^oC 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).

Publications

Species	Application	Title
		Cancer Cell; Adeno-to-squamous transition drives resistance to KRAS inhibition in LKB1 mutant lung cancer; Authors - Xinyuan Tong
	IF	Curr Biol; Gatekeeper function for Short stop at the ring canals of the Drosophila ovary.; Authors - Wen Lu
human	IF	Cell Chem Biol; S1PR3-G12-biased agonist ALESIA targets cancer metabolism and promotes glucose starvation.; Authors - Masayasu Toyomoto
	IF	Elife; A novel mechanism of bulk cytoplasmic transport by cortical dynein in Drosophila ovary.; Authors - Wen Lu
human	IF	Mol Ther Nucleic Acids; Non-targeting control for MISSION shRNA library silences SNRPD3 leading to cell death or permanent growth arrest.; Authors - Maria Czarnek
mouse	IF	Front Cell Dev Biol; Caffeine Induces Autophagy and Apoptosis in Auditory Hair Cells via the SGK1/HIF-1α Pathway.; Authors - Xiaomin Tang

Related Products

Flow Cytometry Perm Buffer (10X)

PF00011-C