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Anti-Tubulin β-3, Polymerized (TUBB3) Antibody (Previously Covance catalog# SMI-62R)

Pricing & Availability
Clone
SMI 62 (See other available formats)
Regulatory Status
RUO
Other Names
CDCBM, CDCBM1, CFEOM3, CFEOM3A, FEOM3, TUBB4, Tubulin beta-3 chain, tubulin beta-III, tubulin beta-4 chain, class III beta-tubulin
Previously
Covance Catalog# SMI-62R
Isotype
Mouse IgG1
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Product Citations
publications
1-SMI-62_polymerized-b-tubulin_purified_IF_022415.jpg
Staining of Polymerized Beta Tubulin (SMI 62) on Hela cells using MeOH fixation.
  • 1-SMI-62_polymerized-b-tubulin_purified_IF_022415.jpg
    Staining of Polymerized Beta Tubulin (SMI 62) on Hela cells using MeOH fixation.
  • 2-SMI-62_Tubulin_beta-3_Polymerized_Antibody_WB_120417
    Total lysates (15 µg protein) from HepG2 and HeLa cells were resolved by electrophoresis (4-20% Tris-glycine gel), transferred to nitrocellulose, and probed with 1:1000 anti-Tubulin β-3, Polymerized (TUBB3) antibody, clone SMI 62. Proteins were visualized using chemiluminescence detection by incubation with HRP Goat anti-Mouse secondary antibody (Cat. No. 405306, 1:3000 dilution). Direct-Blot™ HRP anti-β-actin antibody was used as a loading control (Cat. No. 643807, 1:8000 dilution).
  • 3-SMI-62_Tubulin_beta-3_Polymerized_Antibody_WB_060118
    Whole cell lysates (15 µg protein) from A549 (lane 1) and HeLa (lane 2) cells were resolved by electrophoresis (4-12% Bis-Tris gel), transferred to nitrocellulose, and probed with 0.1 µg/mL (1:5000 dilution) of purified anti-Tubulin Beta 3 (TUBB3) antibody, clone AA10, or anti-Tubulin β-3, Polymerized (TUBB3) antibody (clone SMI 62) (1:1000 dilution). Proteins were visualized using chemiluminescence detection by incubating with 1:3000 dilution of HRP goat anti-mouse-IgG secondary antibody (Cat. No. 405306). Direct-Blot™ HRP anti-β-Actin antibody (1:5000 dilution, Cat. No. 643807) was used as a loading control (lower). Lane M: MW ladder. Cell lysates were loaded in order of decreasing TUBB3 mRNA expression levels; A549 cells express ~14-fold more TUBB3 mRNA than HeLa cells (source: Human Protein Atlas). Lane M: Molecular Weight marker.
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921001 500 µL 279€
 
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Description

Microtubules are required for many well characterized functions in eukaryotic cells, including the movement of chromosomes in mitosis and meiosis, intracellular transport, establishment and maintenance of cellular morphology, cell growth, cell migration, and morphogenesis in multicellular organisms. The building block of a microtubule is the tubulin subunit, a heterodimer of α- and β-tubulin. Both of these monomers are found in all eukaryotes, and their sequences are highly conserved. TUBB3 is primarily expressed in neurons and may be involved in neurogenesis and axon guidance and maintenance.

Product Details
Technical Data Sheet (pdf)

Product Details

Verified Reactivity
Human
Reported Reactivity
Other species
Antibody Type
Monoclonal
Host Species
Mouse
Formulation
Ascites Fluid (contains 0.01M sodium azide).
Preparation
Ascites
Concentration
The concentration is not quantified as this product is sold as undiluted crude mouse ascites fluid. The concentration might vary from lot-to-lot and an estimated concentration would be 1-3 mg/ml.
Storage & Handling
Store at -20°C. Upon initial thawing, apportion into working aliquots and store at -20°C. Avoid repeated freeze-thaw cycles to prevent denaturing the antibody. For long-term storage, keep the antibody at -80°C.
Application

ICC - Quality tested
ELISA, WB
SB - Community verified
Recommended Usage

The optimal working dilution should be determined for each specific assay condition. The extent of permissible dilution of SMI 62 beyond those recommended for general application depends upon nature and concentration of the antigen examined, species of the antigen, method of fixation and kind of section examined.
• WB: 1:1,000
• ICC: 1:1,000
• ELISA: 1:1,000

Application Notes

This antibody is effective in immunoblotting (WB), immunocytochemistry (ICC), and ELISA.

SMI 62 reacts with polymerized ß-tubulin. The antibody has a preference for recognizing polymerized tubulin in immunocytochemistry. This antibody helps reduce background in cell staining applications caused by unpolymerized tubulin in the cytoplasm. Reaction is observed with cold-stabilized tubulin, premitotic, interphase and mitotic nuclei and flagellar tubulin. SMI 62 reacts with most mammalian species in neuronal and non-neuronal tissue. Green algae and molds also react with SMI 62.
 

Observed MW is at 50 kDa.

This clone shows high reactivity towards other ß–tubulin isotypes.

Additional Product Notes

This product has been verified for IHC-F (Immunohistochemistry - frozen tissue sections) on the NanoString GeoMx® Digital Spatial Profiler. The GeoMx® enables researchers to perform spatial analysis of protein and RNA targets in FFPE and fresh frozen human and mouse samples. For more information about our spatial biology products and the GeoMx® platform, please visit our spatial biology page.

Application References

(PubMed link indicates BioLegend citation)
  1. Zhu B, et al. 2010. Am J Physiol Lung Cell Mol Physiol. 299: L493. (WB) PubMed
  2. Fortune B, et al. 2008. Invest. Ophthalmol. Vis. Sci. 49: 255.
Product Citations
  1. Kitakaze K, et al. 2023. Commun Biol. 6:524. PubMed
  2. Zhu B, et al. 2010. Am J Physiol Lung Cell Mol Physiol. 299:L493-501. PubMed
RRID
AB_2565385 (BioLegend Cat. No. 921001)

Antigen Details

Cell Type
Mature Neurons
Biology Area
Cell Biology, Cell Motility/Cytoskeleton/Structure, Neuroscience, Neuroscience Cell Markers
Molecular Family
Microtubules
Gene ID
203068 View all products for this Gene ID
UniProt
View information about Tubulin beta-3 on UniProt.org

Related FAQs

If an antibody clone has been previously successfully used in IBEX in one fluorescent format, will other antibody formats work as well?

It’s likely that other fluorophore conjugates to the same antibody clone will also be compatible with IBEX using the same sample fixation procedure. Ultimately a directly conjugated antibody’s utility in fluorescent imaging and IBEX may be specific to the sample and microscope being used in the experiment. Some antibody clone conjugates may perform better than others due to performance differences in non-specific binding, fluorophore brightness, and other biochemical properties unique to that conjugate.

Will antibodies my lab is already using for fluorescent or chromogenic IHC work in IBEX?

Fundamentally, IBEX as a technique that works much in the same way as single antibody panels or single marker IF/IHC. If you’re already successfully using an antibody clone on a sample of interest, it is likely that clone will have utility in IBEX. It is expected some optimization and testing of different antibody fluorophore conjugates will be required to find a suitable format; however, legacy microscopy techniques like chromogenic IHC on fixed or frozen tissue is an excellent place to start looking for useful antibodies.

Are other fluorophores compatible with IBEX?

Over 18 fluorescent formats have been screened for use in IBEX, however, it is likely that other fluorophores are able to be rapidly bleached in IBEX. If a fluorophore format is already suitable for your imaging platform it can be tested for compatibility in IBEX.

The same antibody works in one tissue type but not another. What is happening?

Differences in tissue properties may impact both the ability of an antibody to bind its target specifically and impact the ability of a specific fluorophore conjugate to overcome the background fluorescent signal in a given tissue. Secondary stains, as well as testing multiple fluorescent conjugates of the same clone, may help to troubleshoot challenging targets or tissues. Using a reference control tissue may also give confidence in the specificity of your staining.

How can I be sure the staining I’m seeing in my tissue is real?

In general, best practices for validating an antibody in traditional chromogenic or fluorescent IHC are applicable to IBEX. Please reference the Nature Methods review on antibody based multiplexed imaging for resources on validating antibodies for IBEX.

Go To Top Version: 5    Revision Date: 01/24/2024

For Research Use Only. Not for diagnostic or therapeutic use.

 

This product is supplied subject to the terms and conditions, including the limited license, located at www.biolegend.com/terms) ("Terms") and may be used only as provided in the Terms. Without limiting the foregoing, BioLegend products may not be used for any Commercial Purpose as defined in the Terms, resold in any form, used in manufacturing, or reverse engineered, sequenced, or otherwise studied or used to learn its design or composition without express written approval of BioLegend. Regardless of the information given in this document, user is solely responsible for determining any license requirements necessary for user’s intended use and assumes all risk and liability arising from use of the product. BioLegend is not responsible for patent infringement or any other risks or liabilities whatsoever resulting from the use of its products.

 

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This data display is provided for general comparisons between formats.
Your actual data may vary due to variations in samples, target cells, instruments and their settings, staining conditions, and other factors.
If you need assistance with selecting the best format contact our expert technical support team.

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