AB263857

Human TUBB3 (beta III Tubulin) knockout HeLa cell lysate

Name: Human TUBB3 (beta III Tubulin) knockout HeLa cell lysate (ab263857) | Abcam
Brand: Abcam Limited
SKU: AB263857
Price: 119000 JPY
Availability: InStock

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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.

別名を表示する

CDCBM, CDCBM1, CFEOM3, CFEOM3A, FEOM3, M(beta)3, M(beta)6, MC1-R, Neuron specific beta III Tubulin, Neuron-specific class III beta-tubulin, QccE-11995, QccE-15186, TBB3_HUMAN, TUBB 4, Tubb 3, Tubulin beta 3, Tubulin beta 4, Tubulin beta-3 chain, Tubulin beta-4 chain, Tubulin beta-III, beta 3 tubulin, beta-4, tuj 1

3 Images

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='/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='/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

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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='/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='/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

細胞タイプ

HeLa

生物種

Human

組織

Cervix

ノックアウト検証方法

Sanger Sequencing,Western blot

ノックアウト変異

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

疾病

Adenocarcinoma

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Reactivity data

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製品の詳細

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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製品内容

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出荷温度及び保存条件

遺伝子名

TUBB3

遺伝子編集のタイプ

Knockout

遺伝子編集の方法

CRISPR technology

ノックアウト検証方法

Sanger Sequencing, Western blot

接合型

Homozygous

出荷温度

Ambient - Can Ship with Ice

短期保存温度

-20°C

長期保存温度

-20°C

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補足情報

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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製品プロトコール

Visit the General protocols
Visit the Troubleshooting

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Abcam product promise

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Human TUBB3 (beta III Tubulin) knockout HeLa cell lysate