Human EHMT1 knockout HCT116 cell line
- Advanced Validation
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EHMT1 KO cell line available to order. KO validated by Next Generation Sequencing, Western blot. Free of charge wild type control provided.
View Alternative Names
DKFZp667M072, EHMT1_HUMAN, Eu-HMTase1, Euchromatic histone methyltransferase 1, Euchromatic histone-lysine N-methyltransferase 1, FLJ12879, FP13812, G9a like protein, G9a-like protein 1, GLP 1, H3-K9-HMTase 5, Histone H3-K9 methyltransferase 5, Histone lysine N methyltransferase H3 lysine 9 specific 5, Histone-lysine N-methyltransferase EHMT1, KIAA1876, Lysine N-methyltransferase 1D, RP11 188C12.1, bA188C12.1
- WB
Lab
Western blot - Human EHMT1 knockout HCT116 cell line (AB287388)
Western blot : Anti-EHMT1 antibody [EPR18667] (ab194299) staining at 1/1000 dilution, shown in green; Mouse anti-CANX [CANX/1543] (ab238078) loading control staining at 1/20000 dilution, shown in magenta. In Western blot, ab194299 was shown to bind specifically to EHMT1. A band was observed at 165 kDa in wild-type HCT 116 cell lysates with no signal observed at this size in EHMT1 knockout cell line. To generate this image, wild-type and EHMT1 knockout HCT 116 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 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.
All lanes:
Western blot - Anti-EHMT2/G9A + EHMT1/GLP antibody [EPR18667] (<a href='/en-us/products/primary-antibodies/ehmt2-g9a-ehmt1-glp-antibody-epr18667-ab194299'>ab194299</a>) at 1/1000 dilution
Lane 1:
Wild-type HCT 116 cell lysate at 20 µg
Lane 2:
EHMT1 knockout HCT 116 cell lysate at 20 µg
Lane 2:
Western blot - Human EHMT1 knockout HCT116 cell line (ab287388)
Secondary
All lanes:
Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution
Observed band size: 165 kDa
false
- NGS
Lab
Next Generation Sequencing - Human EHMT1 knockout HCT116 cell line (AB287388)
164 bp deletion after Thr 53 (allele 1); 127 bp deletion after Thr 53 (allele 2) of WT protein
Reactivity data
Product details
Although we aim to provide customers with a homozygous clone, feasibility will be dependent on the biology of the protein. Should only heterozygous edits be achieved, you will be notified of the outcome and be asked to confirm whether the cell line is acceptable. All clones will be accompanied with DNA sequencing data, and the mutation description.
Recommended control: Human wild-type HCT116 cell line (ab288559). Please note a wild-type cell line is not automatically included with a knockout cell line order, if required please add recommended wild-type cell line at no additional cost using the code WILDTYPE-TMTK1.
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.
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
McCoY5a + 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
EHMT1/GLP plays a critical role in cellular processes such as development and differentiation. It forms a complex with G9a another histone methyltransferase to facilitate these functions. This complex cooperates in controlling gene silencing and the preservation of cellular identity. Such complex mechanisms lead to alterations in gene expression patterns that are essential for normal cellular operation.
Pathways
The methylation activity of EHMT1/GLP participates in the epigenetic regulation pathway and chromatin remodeling pathways. Within these pathways it interacts closely with proteins like HP1 which binds to methylated histones to promote gene silencing. The connections with pathways that influence gene expression affirm its role in maintaining cellular processes like proliferation and embryonic development.
Quality control
STR analysis
CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
Biosafety level
EU: 1 US: 1
Adherent/suspension
Adherent
Gender
Male
Target data
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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