Human CBX4 knockout HEK-293 cell line
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CBX4 KO cell line available to order. KO validated by Next Generation Sequencing. Free of charge wild type control available. Knockout achieved by CRISPR/Cas9; X = 7 bp deletion, 1 bp insertion, 10 bp deletion. To order both knockout and wild-type control cells: select '2 x 1000000 Cells/vial'. To order only knockout cells: select '1000000 Cells/vial'.
View Alternative Names
CBX4_HUMAN, Cbx4 chromobox homolog 4 (Drosophila Pc class), Chromobox homolog 4, Chromobox homolog 4 (Pc class homolog, Drosophila), Chromobox protein homolog 4, E3 SUMO-protein ligase CBX4, NBP 16, NS5ATP1 binding protein 16, PC 2, Pc class 2 homolog, Pc class homolog, Pc class homolog Drosophila, Polycomb 2 homolog, hPc 2
- WB
Lab
Western blot - Human CBX4 knockout HEK-293 cell line (AB261723)
False colour image of Western blot : Anti-CBX4 antibody [EPR23053-7] - ChIP Grade staining at 1/1000 dilution, shown in green; Mouse anti-Alpha Tubulin [DM1A] (ab7291) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab242149 was shown to bind specifically to CBX4. A band was observed at 75 kDa in wild-type HEK-293 cell lysates with no signal observed at this size in Cbx4 knockout cell line ab261723 (knockout cell lysate ab261666). The band observed in the knockout lysate lane below 75 kDa is likely to represent a truncated form of CBX4. This has not been investigated further and the functional properties of the gene product have not been determined. To generate this image, wild-type and Cbx4 knockout HEK-293 cell lysates were analysed.First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3% milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4°C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) at 1/20000 dilution.
All lanes:
Western blot - Anti-CBX4 antibody [EPR23053-7] - ChIP Grade (<a href='/en-us/products/primary-antibodies/cbx4-antibody-epr23053-7-chip-grade-ab242149'>ab242149</a>) at 1/1000 dilution
Lane 1:
Wild-type HEK-293 cell lysate at 20 µg
Lane 2:
Cbx4 CRISPR-Cas9 edited HEK-293 cell lysate at 20 µg
Lane 2:
Western blot - Human CBX4 knockout HEK-293 cell line (ab261723)
Lane 3:
RAW 264.7 cell lysate at 20 µg
Lane 4:
HUVEC cell lysate at 20 µg
Predicted band size: 61 kDa
Observed band size: 75 kDa
false
- NGS
Supplier Data
Next Generation Sequencing - Human CBX4 knockout HEK-293 cell line (AB261723)
Knockout achieved by CRISPR/Cas9; X = 7 bp deletion, 1 bp insertion, 10 bp deletion
Product details
We will provide viable cells that proliferate on revival.
Western blot data indicates that the CRISPR gene edit may have resulted in a truncation of the protein of interest. Please see data images.
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
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
CBX4 operates as a component of the Polycomb Repressive Complex 1 (PRC1) which plays role in controlling gene expression patterns during development. In this complex CBX4 interacts with other core components such as BMI1 and RING1 to facilitate monoubiquitination of histone H2A leading to transcriptional repression. This ability of CBX4 to modulate epigenetic states highlights its importance in critical developmental processes and differentiation.
Pathways
CBX4 significantly influences gene regulatory networks involved in cell cycle and apoptosis pathways. In the cell cycle regulation CBX4 interacts with important proteins such as Cyclin D1 impacting cell proliferation. In apoptosis CBX4 has roles linked to the p53 pathway by modulating transcriptional activity which influences cellular fate. These pathways underline the significance of CBX4 in maintaining balance between cell growth and death.
Quality control
STR analysis
CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
Biosafety level
EU: 2 US: 2
Adherent/suspension
Adherent
Gender
Female
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Product promise
Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.
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