Human TUBB3 (beta III Tubulin) knockout HeLa cell line
- Advanced Validation
- What is this?
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TUBB3 KO cell line available to order. KO validated by Western blot. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, Homozygous: 1 bp insertion in exon 4.
View Alternative Names
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
- WB
Lab
Western blot - Human TUBB3 (beta III Tubulin) knockout HeLa cell line (AB255358)
Lanes 1- 2 : Merged signal (red and green). Green - ab215037 observed at 50 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.
ab215037 was shown to react with beta III tubulin 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 HeLa and TUBB3 knockout HeLa cell lysates 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) overnight at 4° at a 1 in 2000 dilution and a 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 (ab255358)
Predicted band size: 50 kDa
Observed band size: 50 kDa
false
- WB
Lab
Western blot - Human TUBB3 (beta III Tubulin) knockout HeLa cell line (AB255358)
Lanes 1- 2 : Merged signal (red and green). Green - ab52623 observed at 50 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.
ab52623 was shown to react with beta III tubulin 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 HeLa and TUBB3 knockout HeLa cell lysates 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) overnight at 4° at a 1 in 1000 dilution and a 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 (ab255358)
Predicted band size: 50 kDa
Observed band size: 50 kDa
false
- Sanger seq
Unknown
Sanger Sequencing - Human TUBB3 (beta III Tubulin) knockout HeLa cell line (AB255358)
Homozygous : 1 bp insertion in exon 4.
Reactivity data
Product details
Recommended control: Human wild-type HeLa cell line (ab255448). Please note a wild-type cell line is not automatically included with a knockout cell line order, if required please add recommended wild-type cell line at no additional cost using the code WILDTYPE-TMTK1.
We will provide viable cells that proliferate on revival.
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.
What's included?
Properties and storage information
Gene name
Gene editing type
Gene editing method
Knockout validation
Zygosity
Shipped at conditions
Appropriate short-term storage conditions
Appropriate long-term storage conditions
Handling procedures
Initial handling guidelines
Upon arrival, the vial should be stored in liquid nitrogen vapor phase and not at -80°C. Storage at -80°C may result in loss of viability.
1. Thaw the vial in 37°C water bath for approximately 1-2 minutes.
2. Transfer the cell suspension (0.8 mL) to a 15 mL/50 mL conical sterile polypropylene centrifuge tube containing 8.4 mL pre-warmed culture medium, wash vial with an additional 0.8 mL culture medium (total volume 10 mL) to collect remaining cells, and centrifuge at 201 x g (rcf) for 5 minutes at room temperature. 10 mL represents minimum recommended dilution. 20 mL represents maximum recommended dilution.
3. Resuspend the cell pellet in 5 mL pre-warmed culture medium and count using a haemocytometer or alternative cell counting method seed all remaining cells into a T25.
4. Incubate the culture at 37°C incubator with 5% CO2. Check the culture one day after revival and continue to check until 80% confluent. Media change can be given if needed.
5. Once confluent passage into an appropriate flask at a density of 2x104 cells/cm2. Seeding density is given as a guide only and should be scaled to align with individual lab schedules. Cultures should be monitored daily.
Subculture guidelines
- All seeding densities should be based on cell counts gained by established methods.
- A guide seeding density of 2x104 cells/cm2 is recommended.
- Cells should be passaged when they have achieved 80-90% confluence.
Culture medium
DMEM (High Glucose) + 10% FBS
Cryopreservation medium
Cell Freezing Medium-DMSO Serum free media, contains 8.7% DMSO in MEM supplemented with methyl cellulose.
Supplementary information
This supplementary information is collated from multiple sources and compiled automatically.
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.
Quality control
STR analysis
CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
Biosafety level
EU: 2 US: 2
Adherent/suspension
Adherent
Gender
Female
Target data
Complementary Products
Product promise
Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.
For licensing inquiries, please contact partnerships@abcam.com