Human CBX3 (HP1 gamma) knockout HeLa cell line
- Advanced Validation
- What is this?
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CBX3 KO cell line available to order. KO validated by Western blot. Free of charge wild type control available. Knockout achieved by using CRISPR/Cas9, 4 bp deletion in exon 2 and 5 bp deletion in exon 2. To order both knockout and wild-type control cells: select 2 x 1000000Cells/vial. To order only knockout cells: select 1000000Cells/vial.
View Alternative Names
CBX3_HUMAN, Chromobox homolog 3, Chromobox homolog 3 (HP1 gamma homolog, Drosophila), Chromobox protein homolog 3, GAMMA, HECH, HP1 gamma, HP1 gamma homolog, HP1Hs gamma, Heterochromatin like protein 1, Heterochromatin protein 1 homolog gamma, Heterochromatin protein HP1 gamma, Modifier 2 protein
- WB
Lab
Western blot - Human CBX3 (HP1 gamma) knockout HeLa cell line (AB261744)
Lanes 1- 2 : Merged signal (red and green). Green - ab217999 observed at 25 kDa. Red - Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) observed at 50 kDa.
ab217999 was shown to react with HP1 gamma/CBX3 in wild-type HeLa cells in western blot. Loss of signal was observed when knockout cell line ab261744 (knockout cell lysate ab257110) was used. Wild-type HeLa and CBX3 knockout HeLa cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab217999 and Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) overnight at 4°C at a 1 in 2000 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-HP1 gamma/CBX3 antibody [EPR19802] (<a href='/en-us/products/primary-antibodies/hp1-gamma-cbx3-antibody-epr19802-ab217999'>ab217999</a>) at 1/2000 dilution
Lane 1:
Wild-type HeLa cell lysate at 20 µg
Lane 2:
CBX3 knockout HeLa cell lysate at 20 µg
Lane 2:
Western blot - Human CBX3 (HP1 gamma) knockout HeLa cell line (ab261744)
Predicted band size: 20 kDa
Observed band size: 25 kDa
false
- Sanger seq
Unknown
Sanger Sequencing - Human CBX3 (HP1 gamma) knockout HeLa cell line (AB261744)
Allele-2 : 4 bp deletion in exon 2.
- Sanger seq
Unknown
Sanger Sequencing - Human CBX3 (HP1 gamma) knockout HeLa cell line (AB261744)
Allele-1 : 5 bp deletion in exon 2.
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 and ERS Genomics Limited, and is developed with patented technology. For full details of the limited use licenses and relevant 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
HP1 gamma plays a significant role in gene silencing and chromatin structure maintenance. It forms part of a protein complex that includes chromatin remodeling enzymes and transcriptional repressors which help regulate access to genomic DNA. HP1 gamma's association with other chromatin-associated proteins helps modulate the transcriptional response to cellular stimuli ensuring appropriate gene expression patterns needed for normal cell function and development.
Pathways
HP1 gamma is involved in heterochromatin formation and maintenance pathways working alongside other proteins like SUV39H1 and HP1 alpha. Through these pathways HP1 gamma ensures the compact organization of repetitive DNA sequences and suppresses unnecessary transcription. It is also involved in the DNA damage response pathway playing a role in preserving genomic integrity by interacting with proteins such as BRCA1 and 53BP1 involved in DNA repair.
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
Complementary Products
Primary Antibodies
AB217999
Anti-HP1 gamma/CBX3 antibody [EPR19802]
RabMAb|Recombinant|KO Validated
![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HP1 gamma/CBX3 antibody [EPR19802] (AB217999)](https://content.abcam.com/products/images/hp1-gamma-cbx3-antibody-epr19802-ab217999--immunohistochemistry-formalin-pfa-fixed-paraffin-embedded-sections-img74310.jpg)
- 0
- 0
Cell Lines & Lysates
AB255358
Human TUBB3 (beta III Tubulin) knockout HeLa cell line
Advanced Validation
- 0
- 0
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