Human BUB1 knockout HeLa cell line
- Advanced Validation
- What is this?
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- WB
Lab
Western blot - Human BUB1 knockout HeLa cell line (AB265145)
Lanes 1- 2 : Merged signal (red and green). Green - ab195268 observed at 125 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.
ab195268 was shown to react with Bub1 in wild-type HeLa cells in western blot. Loss of signal was observed when knockout cell line ab265145 (knockout cell lysate ab257373) was used. Wild-type HeLa and BUB1 knockout HeLa cell lysates were subjected to SDS-PAGE. ab195268 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated 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-Bub1 antibody [EPR18947] (<a href='/en-us/products/primary-antibodies/bub1-antibody-epr18947-ab195268'>ab195268</a>) at 1/1000 dilution
Lane 1:
Wild-type HeLa cell lysate at 20 µg
Lane 2:
BUB1 knockout HeLa cell lysate at 20 µg
Lane 2:
Western blot - Human BUB1 knockout HeLa cell line (ab265145)
Predicted band size: 122 kDa
Observed band size: 125 kDa
false
- Sanger seq
Unknown
Sanger Sequencing - Human BUB1 knockout HeLa cell line (AB265145)
Homozygous : 1 bp insertion in exon 4.
Complementary Products
Primary Antibodies
AB195268
RabMAb|Recombinant|KO Validated|20ul selling size
![Immunocytochemistry/ Immunofluorescence - Anti-Bub1 antibody [EPR18947] (AB195268)](https://content.abcam.com/products/images/bub1-antibody-epr18947-ab195268--immunocytochemistry-immunofluorescence-img12561.jpg)
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Reactivity data
Product details
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
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.
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
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