- 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='/ko/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 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
보완 제품
Primary Antibodies
AB284808
Anti-FGFR1 alpha (phospho Y654) antibody [EP843(2)] - BSA and Azide free
RabMAb
![Indirect ELISA - Anti-FGFR1 alpha (phospho Y654) antibody [EP843(2)] - BSA and Azide free (AB284808)](https://content.abcam.com/products/images/fgfr1-alpha-phospho-y654-antibody-ep8432-bsa-and-azide-free-ab284808--indirect-elisa-img402544.jpg)
- 0
- 0
Primary Antibodies
AB76464
20ul selling size|RabMAb|Recombinant|KO Validated
![Immunocytochemistry/ Immunofluorescence - Anti-FGFR1 antibody [EPR806Y] (AB76464)](https://content.abcam.com/products/images/fgfr1-antibody-epr806y-ab76464--immunocytochemistry-immunofluorescence-img76790.jpg)
- 5
- 2
제품 세부 정보
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.
제품 구성
특성 및 보관 정보
유전자명
유전자 편집 종류
유전자 편집 방식
Knockout 검증
배송 시 보관 조건
적절한 단기 보관 조건
적절한 장기 보관 조건
추가 정보
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.
품질 관리
STR analysis
CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
Biosafety level
EU: 2 US: 2
Adherent/suspension
Adherent
Gender
Female
초기 취급 지침
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. <p>1. Thaw the vial in 37°C water bath for approximately 1-2 minutes.<br>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.<br>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.<br>4. Incubate the culture at 37°C incubator with 5% CO<sub>2</sub>. Check the culture one day after revival and continue to check until 80% confluent. Media change can be given if needed.<br>5. Once confluent passage into an appropriate flask at a density of 2x10<sup>4</sup> cells/cm<sup>2</sup>. Seeding density is given as a guide only and should be scaled to align with individual lab schedules. Cultures should be monitored daily.</p>
Subculture 가이드라인
- 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.
배양 배지
DMEM (High Glucose) + 10% FBS
동결보존 배지
Cell Freezing Medium-DMSO Serum free media, contains 8.7% DMSO in MEM supplemented with methyl cellulose.
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