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Alexa Fluor® 488 anti-Tubulin β 3 (TUBB3) Antibody (Previously Covance catalog# A488-435L)

Pricing & Availability
Clone
TUJ1 (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# A488-435L
Isotype
Mouse IgG2a, κ
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Product Citations
publications
1_TUJ1_AF488_Tubulin_B_3_Antibody_IHC_011615
CHO cell line expressing Tubulin β 3 stained with Alexa Fluor® 488 anti-Tubulin β 3.
  • 1_TUJ1_AF488_Tubulin_B_3_Antibody_IHC_011615
    CHO cell line expressing Tubulin β 3 stained with Alexa Fluor® 488 anti-Tubulin β 3.
  • 2_TUJ1_A488_Tubulinbeta3_Antibody_IHC-P_2_011718.jpg
    IHC staining of Alexa Fluor® 488 anti-Tubulin β 3 (TUBB3) antibody (clone TUJ1) on formalin-fixed paraffin-embedded human cerebellum tissue. Following antigen retrieval using Sodium Citrate H.I.E.R., the tissue was incubated with 5 µg/ml of the primary antibody for 60 minutes at room temperature. The image was captured with a 40X objective.
  • 3_TUJ1_A488_Tubulinbeta3_Antibody_IHC-P_3_011718.jpg
    IHC staining of Alexa Fluor® 488 anti-Tubulin β 3 (TUBB3) antibody (Clone TUJ1) on formalin-fixed paraffin-embedded normal human cerebellum tissue. Following antigen retrieval using Sodium Citrate H.I.E.R., the tissue was incubated with 1 µg/ml of the primary antibody for 60 minutes at room temperature. The image was captured with a 40X objective.
  • 4_TUJ1_A488_Tubulinbeta3_Antibody_4_101420
    Human lung adenocarcinoma cell line A549 was treated with Fixation Buffer (Cat# 420801) and Permeabilization Wash Buffer (Cat# 421002), and then stained with TUBB3 (clone TUJ1) Alexa Fluor® 488 (filled histogram) or mouse IgG2a, κ Alexa Fluor® 488 isotype control (open histogram).
  • 5_TUJ1_A488_Tubulinbeta3_Antibody_3D_IHC_05112021.png
    Paraformaldehyde-fixed (4%), 500 µm-thick mouse kidney tissue section was processed according to the Ce3DTM Tissue Clearing Kit protocol (cat. no. 427701). The section was costained with anti-Tubulin β 3 (TUBB3) Antibody (clone TUJ1) Alexa Fluor® 488 at 5 µg/mL (green), and anti-mouse CD326 (Ep-CAM) Antibody (clone G8.8) Alexa Fluor® 594 at 5 µg/mL (magenta). The section was then optically cleared and mounted in a sample chamber. The image was captured with a 20X objective using Zeiss 780 confocal microscope and processed by Imaris image analysis software.
    Watch the video.
  • 6_25_Human_Jejunum_BTub3_CD138
    Confocal image of human jejunum sample acquired using the IBEX method of highly multiplexed antibody-based imaging: β-tubulin 3 (yellow) in Cycle 3 and CD138 (purple) in Cycle 5. Tissues were prepared using ~1% (vol/vol) formaldehyde and a detergent. Following fixation, samples are immersed in 30% (wt/vol) sucrose for cryoprotection. Images are courtesy of Drs. Andrea J. Radtke and Ronald N. Germain of the Center for Advanced Tissue Imaging (CAT-I) in the National Institute of Allergy and Infectious Diseases (NIAID, NIH).
Compare all formats See Alexa Fluor® 488 spectral data
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801203 100 µL $352
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Description

Tubulin is the main component of microtubules. In adults, tubulin beta 3 (TUBB3) is primarily expressed in neurons and is commonly used as a neuronal marker. It plays an important role in neuronal cell proliferation and differentiation. Mutations in this gene cause congenital fibrosis of the type 3 extraocular muscles. Tubulin beta 3 (TUBB3) is also found in a wide range of tumors. Studies indicate that it is a predictive and prognostic marker in various tumors.

Product Details
Technical data sheet

Product Details

Verified Reactivity
Human, Mouse, Rat
Antibody Type
Monoclonal
Host Species
Mouse
Immunogen
This antibody was raised against microtubules derived from rat brain.
Formulation
Alexa Fluor® Labeled Antibody (in PBS + 50% glycerol + 0.03% thimerosal).
Concentration
1.0 mg/ml
Storage & Handling
The antibody solution should be stored undiluted between 2°C and 8°C, and protected from prolonged exposure to light. Do not freeze.
Application

ICC - Quality tested
IHC-P, ICFC, 3D IHC - Verified
SB - Reported in the literature, not verified in house
Recommended Usage

Each lot of this antibody is quality control tested by immunocytochemistry. For immunocytochemistry, a dilution of 1:500 is recommended. For immunohistochemistry, a concentration range of 1.0 - 5.0 µg/ml is suggested. For flow cytometric staining, the suggested use of this reagent is 5 µL per million cells in 100 µL staining volume or 5 µL per 100 µL of whole blood. For 3D immunohistochemistry on formalin-fixed tissues, a concentration of 5.0 µg/mL is suggested. It is recommended that the reagent be titrated for optimal performance for each application.


Alexa Fluor® and Pacific Blue™ are trademarks of Life Technologies Corporation.

View full statement regarding label licenses
Excitation Laser
Blue Laser (488 nm)
Application Notes

Additional reported applications (for the relevant formats) include: flow cytometry4, immunofluorescence microscopy1-5,7, immunohistochemistry5,7, Western blotting8, and spatial biology (IBEX)9,10.

This antibody is well characterized and highly reactive to neuron specific Class III ß-tubulin (ßIII). TUJ1 does not identify ß-tubulin found in glial cells. TUJ1 recognizes an epitope located within the last 15 C-terminal residues8.

Additional Product Notes

Iterative Bleaching Extended multi-pleXity (IBEX) is a fluorescent imaging technique capable of highly-multiplexed spatial analysis. The method relies on cyclical bleaching of panels of fluorescent antibodies in order to image and analyze many markers over multiple cycles of staining, imaging, and, bleaching. It is a community-developed open-access method developed by the Center for Advanced Tissue Imaging (CAT-I) in the National Institute of Allergy and Infectious Diseases (NIAID, NIH).

Application References

(PubMed link indicates BioLegend citation)
  1. Nishimura K, et al. 2017. PLoS One. 12(1): e0170568. (ICC)
  2. Jongbloets J, et al. 2017. Nat Commun. 8: 14666. (ICC) PubMed
  3. Liu W.J, et al. 2015. Eur J Histochem. 59(1): 2464. (ICC) PubMed
  4. Chintalapudi SR, et al. 2016. Front Aging Neursci. 8:93. (FC, ICC) PubMed
  5. Ambasudhan R, et al. 2011. Cell Stem Cell. 9(2):113. (IHC, ICC)
  6. Hu X., et al. 2006. Nature Neuroscicene. 9(12):1520. (WB) PubMed
  7. Zechner D., et al. 2003. Develop Biology. 258(2):406. (ICC, IHC)
  8. Lee MK, et al. 1990. Proc. Natl. Acad. Sci. USA 18:7195. (WB)
  9. Radtke AJ, et al. 2020. Proc Natl Acad Sci USA. 117:33455-33465. (SB) PubMed
  10. Radtke AJ, et al. 2022. Nat Protoc. 17:378-401. (SB) PubMed
Product Citations
  1. Saraswathula A, et al. 2023. Int Forum Allergy Rhinol. 13:230. PubMed
  2. Tokutake K, et al. 2022. Int J Mol Sci. 23: . PubMed
  3. Roque CG, et al. 2023. Sci Adv. 9:eadd2671. PubMed
  4. Vereertbrugghen A, et al. 2023. J Neuroinflammation. 20:120. PubMed
  5. Ruan H, et al. 2023. Acta Pharm Sin B. 13:2202. PubMed
  6. Babetto E, et al. 2020. Nat Neurosci. 23:1215. PubMed
  7. Lu R, et al. 2021. Int J Mol Sci. 22:00. PubMed
  8. Giacomini C, et al. 2016. Mol Biol Cell. 27: 35 - 47. PubMed
  9. McCurdy EP et al. 2019. Cell Rep. 29(2):363-377 . PubMed
  10. Guzmán M, et al. 2020. Immunology. 161:148. PubMed
  11. Rocktäschel P, et al. 2019. Epilepsy Behav. 101:106581. PubMed
  12. Kim Y, et al. 2016. Sci Rep. 6: 23799. PubMed
  13. Inlender T, et al. 2021. Scientific Reports. 11(1):12796. PubMed
  14. Arsić A, et al. 2020. Sci Rep. 10:6441. PubMed
  15. Wang S, et al. 2022. PLoS Pathog. 18:e1010281. PubMed
  16. Stachtea X, et al. 2015. PLoS One. 10: e0140279. PubMed
  17. Larhammar M et al. 2017. eLife. 6 pii: e20725. PubMed
  18. Jalilian E, et al. 2022. Stem Cell Res Ther. 13:425. PubMed
  19. Chen M, et al. 2019. Cell Stem Cell. 25:501. PubMed
  20. Martínez JC et al. 2019. Neuron. 104(5):931-946 . PubMed
  21. Geisler S, et al. 2019. JCI Insight. 4:e129920. PubMed
  22. Žiak J, et al. 2020. EMBO Rep. e48512:21. PubMed
  23. Chung KM, et al. 2022. Cell Rep. 41:111488. PubMed
RRID
AB_2564757 (BioLegend Cat. No. 801203)
Disclaimer

Covered by US patents 5,675,063 and 7,429,487. Sold under license from Epitomics.

Antigen Details

Structure
Tubulin β 3 is a 450 amino acid protein with a molecular mass of ~50 kD.
Distribution

Tissue distribution: central and peripheral nervous system.
Cellular distribution: cytosol, cytoskeleton and nucleus.

Function
Tubulin β 3 is the major constituent of microtubules, and plays a critical role in proper axon guidance and maintenance.
Interaction
Alpha tubulin, kinesin and dynein.
Cell Type
Mature Neurons, Neural Stem Cells
Biology Area
Cell Biology, Neuroscience, Neuroscience Cell Markers, Stem Cells
Molecular Family
Microtubules
Antigen References

1. Zhao X, et al. 2017. Med Sci Monit. 22: 3915.
2. Lebok P, et al. 2016. Oncol Lett. 11(3):1987.
3. Du J, et al. 2015. BMC Cancer. 15:536. PubMed
4. Rogue DM., et al. 2013. Clin Exp Metastasis. 31(1): 101.
5. Ploussard G, et al. 2010. Cancer Res. 70(22):9253. PubMed

Gene ID
10381 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: 6    Revision Date: 04/26/2022

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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