Human EHMT2 (G9A) knockout HeLa cell line
- Advanced Validation
- What is this?
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(0 Publication)
- WB
Lab
Western blot - Human EHMT2 (G9A) knockout HeLa cell line (AB265149)
Lanes 1-4 : Merged signal (red and green). Green - ab133482 observed at 160 kDa. Red - loading control ab8245 observed at 37 kDa.
ab133482 Anti-EHMT2/G9A antibody [EPR4019(2)] was shown to specifically react with EHMT2/G9A in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab265149 (knockout cell lysate ab257080) was used. Wild-type and EHMT2/G9A knockout samples were subjected to SDS-PAGE. ab133482 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4° at 1 in 1000 dilution 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-EHMT2/G9A antibody [EPR4019(2)] - ChIP Grade (<a href='/en-us/products/primary-antibodies/ehmt2-g9a-antibody-epr40192-chip-grade-ab133482'>ab133482</a>) at 1/1000 dilution
Lane 1:
Wild-type HeLa cell lysate at 20 µg
Lane 2:
EHMT2/G9A knockout HeLa cell lysate at 20 µg
Lane 2:
Western blot - Human EHMT2 (G9A) knockout HeLa cell line (ab265149)
Lane 3:
HEK-293 cell lysate at 20 µg
Lane 4:
HepG2 cell lysate at 20 µg
Secondary
All lanes:
Western blot - Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-irdye-800cw-preadsorbed-ab216773'>ab216773</a>) at 1/20000 dilution
Predicted band size: 132 kDa
Observed band size: 160 kDa
false
- WB
Lab
Western blot - Human EHMT2 (G9A) knockout HeLa cell line (AB265149)
Lanes 1-4 : Merged signal (red and green). Green - ab185050 observed at 160 kDa. Red - loading control ab8245 observed at 37 kDa.
ab185050 Anti-EHMT2/G9A antibody [EPR18894] was shown to specifically react with EHMT2/G9A in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab265149 (knockout cell lysate ab257080) was used. Wild-type and EHMT2/G9A knockout samples were subjected to SDS-PAGE. ab185050 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4° at 1 in 1000 dilution 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-EHMT2/G9A antibody [EPR18894] - ChIP Grade (<a href='/en-us/products/primary-antibodies/ehmt2-g9a-antibody-epr18894-chip-grade-ab185050'>ab185050</a>) at 1/1000 dilution
Lane 1:
Wild-type HeLa cell lysate at 20 µg
Lane 2:
EHMT2/G9A knockout HeLa cell lysate at 20 µg
Lane 2:
Western blot - Human EHMT2 (G9A) knockout HeLa cell line (ab265149)
Lane 3:
HEK-293 cell lysate at 20 µg
Lane 4:
HepG2 cell lysate at 20 µg
Secondary
All lanes:
Western blot - Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-irdye-800cw-preadsorbed-ab216773'>ab216773</a>) at 1/20000 dilution
Predicted band size: 132 kDa
Observed band size: 160 kDa
false
- Cell Culture
Unknown
Cell Culture - Human EHMT2 (G9A) knockout HeLa cell line (AB265149)
Representative images of EHMT2 knockout HeLa cells, low and high confluency examples (top left and right respectively) and wild-type HeLa cells, low and high confluency (bottom left and right respectively) showing typical adherent, epithelial-like morphology. Images were captured at 10X magnification using a EVOS XL Core microscope.
- Sanger seq
Unknown
Sanger Sequencing - Human EHMT2 (G9A) knockout HeLa cell line (AB265149)
Allele-2 : 370 bp deletion in exon 7.
- Sanger seq
Unknown
Sanger Sequencing - Human EHMT2 (G9A) knockout HeLa cell line (AB265149)
Allele-1 : 2 bp deletion in exon 7.
Complementary Products
Primary Antibodies
AB185050
Anti-EHMT2/G9A antibody [EPR18894] - ChIP Grade
RabMAb|Advanced Validation|Recombinant|KO Validated
![Immunocytochemistry/ Immunofluorescence - Anti-EHMT2/G9A antibody [EPR18894] - ChIP Grade (AB185050)](https://content.abcam.com/products/images/ehmt2-g9a-antibody-epr18894-chip-grade-ab185050--immunocytochemistry-immunofluorescence-img14082.jpg)
- 5
- 4
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
EHMT2/G9a regulates gene expression by altering chromatin accessibility. It is part of a chromatin remodeling complex interacting with proteins such as ANMTA02. Through its methyltransferase activity it maintains an epigenetic silence state important for embryonic development differentiation and cellular identity. EHMT2/G9a plays a significant role in the repression of transcriptional activity of tumor suppressor genes and developmental genes.
Pathways
G9a functions in epigenetic signaling pathways and influences processes like the cell cycle and DNA damage repair. It interacts with proteins in the Polycomb repressive complex such as EZH2 to regulate gene silencing. Additionally in the DNA damage response pathway G9a can associate with histone deacetylases to stabilize chromatin and prevent unwarranted gene expression.
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