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Purified anti-Tubulin β 3 (TUBB3) Antibody (Previously Covance catalog# MMS-435P)

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# MMS-435P
Isotype
Mouse IgG2a, κ
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Product Citations
publications
A_TUJ1_PURE_Tubulin-beta3_IHCP_Hu_011618
IHC staining of purified anti-Tubulin β 3 (TUBB3) antibody (clone TUJ1) on formalin-fixed paraffin-embedded human brain tissue. Following antigen retrieval using Sodium Citrate H.I.E.R., the tissue was incubated with 1 µg/mL of the primary antibody overnight at 4°C. BioLegend’s Ultra Streptavidin (USA) HRP Detection Kit (Multi-Species, DAB, Cat. No. 929901) was used for detection followed by hematoxylin counterstaining, according to the protocol provided. The image was captured with a 40X objective.
  • A_TUJ1_PURE_Tubulin-beta3_IHCP_Hu_011618
    IHC staining of purified anti-Tubulin β 3 (TUBB3) antibody (clone TUJ1) on formalin-fixed paraffin-embedded human brain tissue. Following antigen retrieval using Sodium Citrate H.I.E.R., the tissue was incubated with 1 µg/mL of the primary antibody overnight at 4°C. BioLegend’s Ultra Streptavidin (USA) HRP Detection Kit (Multi-Species, DAB, Cat. No. 929901) was used for detection followed by hematoxylin counterstaining, according to the protocol provided. The image was captured with a 40X objective.
  • B_TUJ1_PURE_Tubulin-beta3_IHCP_Rat_011618
    IHC staining of purified anti-Tubulin β 3 (TUBB3) antibody (clone TUJ1) on formalin-fixed paraffin-embedded rat brain tissue. Following antigen retrieval using Sodium Citrate H.I.E.R., the tissue was incubated with 0.5 µg/mL of the primary antibody overnight at 4°C. BioLegend’s Ultra Streptavidin (USA) HRP Detection Kit (Multi-Species, DAB, Cat. No. 929901) was used for detection followed by hematoxylin counterstaining, according to the protocol provided. The image was captured with a 40X objective.
  • C_TUJ1_PURE_Tubulin-beta3_WB_011618
    Western blot of purified anti-Tubulin β 3 (TUBB3) antibody (clone TUJ1). Lane 1: Molecular weight marker; Lane 2: 20 µg of human brain lysate; Lane 3: 20 µg of Mouse brain lysate; Lane 4: 20 µg of rat brain lysate. The blots were incubated with 1 ug/mL of TUJ1 or mouse IgG overnight at 4°C, followed by incubation with HRP-labeled goat anti-mouse IgG (Cat. No. 405306). Enhanced chemiluminescence was used as the detection system.
  • D_TUJ1_PURE_Tubulin-beta3_ICC_011618
    ICC staining of purified anti-Tubulin β 3 (TUBB3) antibody (clone TUJ1) on SH-SY5Y neuroblastoma cells. The cells were fixed by 4% PFA, permeabilized with a buffer containing 0.1% Triton X-100 and 0.25% BSA, and blocked with 2% Normal Goat Serum and 0.02% BSA. The cells were then incubated with 1 µg/mL of the primary antibody for 3 hours at room temperature, followed by incubation with 0.5 µg/mL Alexa Fluor® 594 goat anti-mouse IgG (Cat. No. 405326) for 1 hour at room temperature. Nuclei were counterstained with DAPI. The image was captured with a 60X objective Scale bar: 20 µm
  • E_Ankita_Patil
    Methanol fixed superior cervical ganglion neurons were stained with anti-tubulin β3 (pink) and anti-TRIM46 (aqua), then stained with secondary antibodies, phalloidin (orange) and DAPI (blue). Image generously submitted to the 2017 Cell Life Imaging Competition by Ankita Patil from Drexel University.
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801213 25 µL 85€
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801201 100 µL 212€
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801202 250 µL 313€
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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 (pdf)

Product Details

Reactivity
Human,Mouse,Rat
Antibody Type
Monoclonal
Host Species
Mouse
Immunogen
This antibody was raised against microtubules derived from rat brain.
Formulation
Phosphate-buffered solution + 0.03% Thimerosal.
Preparation
The antibody was purified by affinity chromatography.
Concentration
1.0 mg/ml
Storage & Handling
The antibody solution should be stored undiluted between 2°C and 8°C. Please note the storage condition for this antibody has been changed from -20°C to between 2°C and 8°C. You can also check your vial or your CoA to find the most accurate storage condition for this antibody.
Application

IHC-P - Quality tested
WB, ICC - Verified
FC - Reported in the literature, not verified in house
SB - Community verified
Recommended Usage

Each lot of this antibody is quality control tested by formalin-fixed paraffin-embedded immunohistochemical staining. For immunohistochemistry, a concentration range of 1.0 - 5.0 µg/ml is suggested. For Western blotting, the suggested use of this reagent is 1.0 - 5.0 µg/ml. For immunocytochemistry, a concentration range of 1.0 - 5.0 μg/ml is recommended. It is recommended that the reagent be titrated for optimal performance for each application.

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

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)
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  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
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RRID
AB_2313773 (BioLegend Cat. No. 801213)
AB_2313773 (BioLegend Cat. No. 801201)
AB_2313773 (BioLegend Cat. No. 801202)
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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.

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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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