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BUB1 KO cell line available now. KO validated by Western blot. Free of charge wild type control available. Knockout achieved by using CRISPR/Cas9, Homozygous: 1 bp insertion in exon 4.

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Images

Western blot - Human BUB1 knockout HeLa cell line (AB265145), expandable thumbnail
  • Sanger Sequencing - Human BUB1 knockout HeLa cell line (AB265145), expandable thumbnail

Key facts

Cell type

HeLa

Species or organism

Human

Tissue

Cervix

Form

Liquid

Knockout validation

Sanger Sequencing, Western blot

Mutation description

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

Alternative names

BUB1 budding uninhibited by benzimidazoles 1 homolog, BUB1 budding uninhibited by benzimidazoles 1 homolog (yeast), BUB1 mitotic checkpoint serine/threonine kinase, BUB1, S. cerevisiae, homolog of, BUB1A, BUB1L, BUB1_HUMAN, Budding uninhibited by benzimidazoles 1 (yeast homolog), Budding uninhibited by benzimidazoles 1 homolog, Budding uninhibited by benzimidazoles 1, S. cerevisiae, homolog of, Homolog of mitotic checkpoint gene BUB1, Mitotic checkpoint gene BUB1, Mitotic checkpoint serine/threonine-protein kinase BUB1, Mitotic spindle checkpoint kinase, Putative serine/threonine protein kinase, hBUB1

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BUB1 KO cell line available now. KO validated by Western blot. Free of charge wild type control available. Knockout achieved by using CRISPR/Cas9, Homozygous: 1 bp insertion in exon 4.

Alternative names

BUB1 budding uninhibited by benzimidazoles 1 homolog, BUB1 budding uninhibited by benzimidazoles 1 homolog (yeast), BUB1 mitotic checkpoint serine/threonine kinase, BUB1, S. cerevisiae, homolog of, BUB1A, BUB1L, BUB1_HUMAN, Budding uninhibited by benzimidazoles 1 (yeast homolog), Budding uninhibited by benzimidazoles 1 homolog, Budding uninhibited by benzimidazoles 1, S. cerevisiae, homolog of, Homolog of mitotic checkpoint gene BUB1, Mitotic checkpoint gene BUB1, Mitotic checkpoint serine/threonine-protein kinase BUB1, Mitotic spindle checkpoint kinase, Putative serine/threonine protein kinase, hBUB1

Key facts

Cell type

HeLa

Form

Liquid

Mutation description

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

Antibiotic resistance

Puromycin 1µg/mL

Disease

Adenocarcinoma

Concentration
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Properties

Gene name

BUB1

Gene editing type

Knockout

Gene editing method

CRISPR technology

Knockout validation

Sanger Sequencing, Western blot

Zygosity

Homozygous

Quality control

STR analysis

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

Cell culture

Biosafety level

EU: 2 US: 2

Adherent/suspension

Adherent

Gender

Female

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.

Storage

Shipped at conditions

Dry Ice

Appropriate long-term storage conditions

-196°C

Notes

Recommended control: Human wild-type HeLa cell line (Human wild-type HeLa cell line ab255928). 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.

Supplementary info

Activity summary

Bub1 protein often referred to as 'Bub1' plays a mechanical role in cell cycle regulation especially during mitosis. It is known by other names such as BUB1 mitotic checkpoint serine/threonine kinase. Bub1 has a molecular mass of approximately 120 kDa. This protein is expressed in various tissues with higher levels in rapidly dividing cells. The location of its action is mainly the nucleus where it associates with kinetochores during cell division functioning in the spindle assembly checkpoint.

Biological function summary

Bub1 protein acts as a serine/threonine-protein kinase that is essential for proper chromosome segregation. It forms part of the spindle checkpoint complex along with other proteins such as BubR1 and Mad2. This complex ensures that chromosomes are correctly attached to the spindle microtubules before anaphase begins. Bub1's kinase activity contributes to the prevention of premature anaphase onset thereby helping maintain genomic stability.

Pathways

Bub1 is significantly involved in the mitotic checkpoint pathway and the cell cycle control pathway. Within these pathways Bub1 interacts closely with proteins like CDC20 and APC/C which are integral parts of the anaphase-promoting complex. Bub1 modifies the activity of these proteins to delay anaphase onset until all chromosomes are properly aligned ensuring the fidelity of cell division.

Associated diseases and disorders

Bub1 protein shows a connection to various cancer types notably breast cancer and colorectal cancer. Defects or overexpression in Bub1 can disrupt the spindle assembly checkpoint leading to chromosome missegregation and aneuploidy hallmarks of cancerous cells. Bub1 is also linked to other spindle checkpoint proteins such as Mad1 and Bub3. Malfunction in any of these proteins including Bub1 can contribute to tumorigenesis due to impaired cell cycle control.

Product promise

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In the unlikely event of one of our products not working as expected, you are covered by our product promise.

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Product protocols

For this product, it's our understanding that no specific protocols are required. You can:

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

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