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AB263857

Human TUBB3 (beta III Tubulin) knockout HeLa cell lysate

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TUBB3 KO cell lysate available now. KO validated by Western blot. Free of charge wild type control included. Knockout achieved by using CRISPR/Cas9, Homozygous: 1 bp insertion in exon 4.
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Western blot - Human TUBB3 (beta III Tubulin) knockout HeLa cell lysate (AB263857)
  • WB

Lab

Western blot - Human TUBB3 (beta III Tubulin) knockout HeLa cell lysate (AB263857)

Lane 1 : Wild-type HeLa cell lysate (20µg)

Lane 2 : TUBB3 knockout HeLa cell lysate (20µg)

Lanes 1- 2 : Merged signal (red and green). Green - ab52623 observed at 50 kDa. Red - loading control ab8245 observed at 37 kDa.

ab52623 Recombinant Anti-beta III Tubulin antibody [EP1569Y] was shown to specifically react with TUBB3 in wild-type HeLa cells in western blot. Loss of signal was observed when knockout cell line ab255358 (knockout cell lysate ab263857) was used. Wild-type and TUBB3 knockout samples were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab52623 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-beta III Tubulin antibody [EP1569Y] - Neuronal Marker (<a href='/en-us/products/primary-antibodies/beta-iii-tubulin-antibody-ep1569y-neuronal-marker-ab52623'>ab52623</a>) at 1/1000 dilution

Lane 1:

Wild-type HeLa cell lysate at 20 µg

Lane 2:

TUBB3 knockout HeLa cell lysate at 20 µg

Lane 2:

Western blot - Human TUBB3 (beta III Tubulin) knockout HeLa cell line (<a href='/en-us/products/cell-lines/human-tubb3-beta-iii-tubulin-knockout-hela-cell-line-ab255358'>ab255358</a>)

Predicted band size: 50 kDa

Observed band size: 50 kDa

false

Western blot - Human TUBB3 (beta III Tubulin) knockout HeLa cell lysate (AB263857)
  • WB

Lab

Western blot - Human TUBB3 (beta III Tubulin) knockout HeLa cell lysate (AB263857)

Lane 1 : Wild-type HeLa cell lysate (20µg)

Lane 2 : TUBB3 knockout HeLa cell lysate (20µg)

Lanes 1- 2 : Merged signal (red and green). Green - ab215037 observed at 50 kDa. Red - loading control ab8245 observed at 37 kDa.

ab215037 Recombinant Anti-beta III Tubulin antibody [EPR19591] was shown to specifically react with TUBB3 in wild-type HeLa cells in western blot. Loss of signal was observed when knockout cell line ab255358 (knockout cell lysate ab263857) was used. Wild-type and TUBB3 knockout samples were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab215037 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 2000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-beta III Tubulin antibody [EPR19591] - Neuronal Marker (<a href='/en-us/products/primary-antibodies/beta-iii-tubulin-antibody-epr19591-neuronal-marker-ab215037'>ab215037</a>) at 1/2000 dilution

Lane 1:

Wild-type HeLa cell lysate at 20 µg

Lane 2:

TUBB3 knockout HeLa cell lysate at 20 µg

Lane 2:

Western blot - Human TUBB3 (beta III Tubulin) knockout HeLa cell line (<a href='/en-us/products/cell-lines/human-tubb3-beta-iii-tubulin-knockout-hela-cell-line-ab255358'>ab255358</a>)

Predicted band size: 50 kDa

Observed band size: 50 kDa

false

Sanger Sequencing - Human TUBB3 (beta III Tubulin) knockout HeLa cell lysate (AB263857)
  • Sanger seq

Unknown

Sanger Sequencing - Human TUBB3 (beta III Tubulin) knockout HeLa cell lysate (AB263857)

Homozygous : 1 bp insertion in exon 4

Key facts

Cell type

HeLa

Species or organism

Human

Tissue

Cervix

Knockout validation

Sanger Sequencing,Western blot

Mutation description

Knockout achieved by using CRISPR/Cas9, Homozygous: 1 bp insertion in exon 4.

Disease

Adenocarcinoma

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Complementary Products

Cell Lines & Lysates

AB255358

Human TUBB3 (beta III Tubulin) knockout HeLa cell line

Advanced Validation

Western blot - Human TUBB3 (beta III Tubulin) knockout HeLa cell line (AB255358)

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Primary Antibodies

AB18207

Anti-beta III Tubulin antibody - Neuronal Marker

BOND RX™ Validated|Lab Essentials|KO Validated

Immunocytochemistry/ Immunofluorescence - Anti-beta III Tubulin antibody - Neuronal Marker (AB18207)

  • 5
  • 67
Proteins & Peptides

AB259412

Recombinant human VEGF 165A protein (Active)

Premium Bioactive

Mass Spectrometry - Recombinant human VEGF 165A protein (Active) (AB259412)

  • 0
  • 0
Assay Kits

AB235644

Human Beta-III Tubulin ELISA Kit

Recombinant|SimpleStep

Sandwich ELISA - Human Beta-III Tubulin ELISA Kit (AB235644)

  • 0
  • 0
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Reactivity data

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Product details

Knockout cell lysate achieved by CRISPR/Cas9.

REACH authorisation
Abcam has not and does not intend to apply for the REACH Authorisation of customers' uses of products that contain European Authorisation list (Annex XIV) substances.
It is the responsibility of our customers to check the necessity of application of REACH Authorisation, and any other relevant authorisations, for their intended uses.

Lysate preparation: Our lysates are made using RIPA buffer to which we add a protease inhibitor cocktail and phosphatase inhibitor cocktail (ratio: 300:100:10). This means that the protein of interest is denatured. If you require a native form of the protein please use the live cell version. Please refer to our lysis protocol for further details on how our lysates are prepared.

User storage instructions: Lyophilizate may be stored at 4°C. After reconstitution, store at -20°C for short-term storage or -80°C for long-term storage.

This product is subject to limited use licenses from The Broad Institute, ERS Genomics Limited and Sigma-Aldrich Co. LLC, and is developed with patented technology. For full details of the licenses and patents please refer to our limited use license and patent pages.

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What's included?

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Properties and storage information

Gene name
TUBB3
Gene editing type
Knockout
Gene editing method
CRISPR technology
Knockout validation
Sanger Sequencing, Western blot
Zygosity
Homozygous
Shipped at conditions
Ambient - Can Ship with Ice
Appropriate short-term storage conditions
-20°C
Appropriate long-term storage conditions
-20°C
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Supplementary information

This supplementary information is collated from multiple sources and compiled automatically.

Beta III tubulin often referred to as βIII tubulin or beta-tubulin 3 is a microtubule element involved in the cellular cytoskeleton structure. This protein with a molecular mass of approximately 55 kDa plays an important role in the development and maintenance of neuron structures. It is most prominently expressed in neurons within the central and peripheral nervous systems. In these cells beta III tubulin contributes to microtubule polymerization facilitating the formation of the intricate network necessary for cell shape intracellular transport and division.
Biological function summary

Beta III tubulin acts in stabilizing microtubules which are essential for proper neuronal function. It exists as part of the tubulin dimer complex partnering with alpha-tubulin to form the building blocks of microtubules. This particular isotype is often used as a marker for neuronal differentiation due to its specific expression pattern in neural tissues. Its regulation and expression levels are important for neurogenesis and maintaining neuronal plasticity.

Pathways

Beta III tubulin plays a role in neuron-specific intracellular transport and signaling pathways like the Rho GTPase and MAPK signaling pathways. These pathways are involved in cell cycle regulation and apoptotic processes. Relationships with other proteins such as kinesins and dyneins are important in these pathways influencing intracellular transport and signaling through binding and moving along the microtubule tracks created by beta-tubulin isotypes including beta III tubulin.

Abnormal beta III tubulin expression has been associated with cancer particularly in tumors originating from neuronal lineage such as glioblastomas. Overexpression or mutations can contribute to chemoresistance complicating treatment for certain types of cancer. Beta III tubulin is also linked to neurodevelopmental disorders as it affects proper neural network formation and stability. Its relationship with tumor protein p53 is noted in cancer pathways as p53 can influence beta III tubulin expression impacting cellular proliferation and apoptosis in oncogenic processes.
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Quality control

STR analysis

CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX

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Cell culture

Biosafety level

EU: 2 US: 2

Adherent/suspension

Adherent

Gender

Female

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Product protocols

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Product promise

We are committed to supporting your work with high-quality reagents, and we're here for you every step of the way. In the unlikely event that one of our products does not perform as expected, you're protected by our Product Promise.
For full details, please see our Terms & Conditions

Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.

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