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AB257373

Human BUB1 knockout HeLa cell lysate

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BUB1 KO cell lysate available now. KO validated by Western blot. Free of charge wild type control included. Knockout achieved by using CRISPR/Cas9, 1 bp insertion in exon4.
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Western blot - Human BUB1 knockout HeLa cell lysate (AB257373)
  • WB

Lab

Western blot - Human BUB1 knockout HeLa cell lysate (AB257373)

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

Lane 2 : BUB1 knockout HeLa cell lysate (20µg)

Lanes 1- 2 : Merged signal (red and green). Green - ab195268 observed at 125 kDa. Red - loading control, ab8245 observed at 37 kDa.

ab195268 Anti-Bub1 antibody [EPR18947] was shown to specifically 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 and Bub1 knockout samples were subjected to SDS-PAGE. ab195268 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 1000 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-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 (<a href='/en-us/products/cell-lines/human-bub1-knockout-hela-cell-line-ab265145'>ab265145</a>)

Predicted band size: 122 kDa

Observed band size: 125 kDa

false

Sanger Sequencing - Human BUB1 knockout HeLa cell lysate (AB257373)
  • Sanger seq

Unknown

Sanger Sequencing - Human BUB1 knockout HeLa cell lysate (AB257373)

Homozygous : 1 bp insertion in exon4

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, 1 bp insertion in exon4.

Disease

Adenocarcinoma

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Complementary Products

Primary Antibodies

AB195268

Anti-Bub1 antibody [EPR18947]

RabMAb|Recombinant|KO Validated|20ul selling size

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AB127642

Recombinant Human Bub1 protein

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AB288799

Recombinant Human VCAM1 protein (Active)

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Reactivity data

{ "title": "Reactivity Data", "filters": { "stats": ["", "Reactivity", "Dilution Info", "Notes"] }, "values": { "WB": { "reactivity":"TESTED_AND_REACTS", "dilution-info":"", "notes":"<p></p>" } } }
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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
BUB1
Gene editing type
Knockout
Gene editing method
CRISPR technology
Knockout validation
Sanger Sequencing, Western blot
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.

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.

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.
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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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