Human DGCR8 knockout HCT116 cell line
- Advanced Validation
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DGCR8 KO cell line available to order. KO validated by Next Generation Sequencing, Western blot. Free of charge wild type control provided.
View Alternative Names
C22orf12, D16H22S788E, D16Wis2, DGCR8 microprocessor complex subunit, DGCR8_HUMAN, DGCRK 6, DiGeorge syndrome critical region 8, DiGeorge syndrome critical region gene 8, Gy1, Microprocessor complex subunit DGCR8, pasha
- WB
Lab
Western blot - Human DGCR8 knockout HCT116 cell line (AB287366)
Western blot : Anti-DGCR8 antibody [EPR18757] (ab191875) staining at 1/1000 dilution, shown in green; Mouse anti-Alpha Tubulin [DM1A] (ab7291) loading control staining at 1/20000 dilution, shown in magenta. In Western blot, ab191875 was shown to bind specifically to DGCR8. A band was observed at 100 kDa in wild-type HCT 116 cell lysates with no signal observed at this size in DGCR8 knockout cell line. To generate this image, wild-type and DGCR8 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 5 % 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-DGCR8 antibody [EPR18757] (<a href='/en-us/products/primary-antibodies/dgcr8-antibody-epr18757-ab191875'>ab191875</a>) at 1/1000 dilution
Lane 1:
Wild-type HCT 116 cell lysate at 20 µg
Lane 2:
Western blot - Human DGCR8 knockout HCT116 cell line (ab287366)
Lane 2:
DGCR8 knockout HCT 116 cell lysate at 20 µg
Lane 3:
Wild-type A549 ab288558 cell lysate at 20 µg
Lane 4:
DGCR8 knockout A549 <a href='/en-us/products/cell-lines/human-dgcr8-knockout-a549-cell-line-ab287368'>ab287368</a> cell lysate at 20 µg
Secondary
Lanes 1 - 4:
Goat anti-Rabbit IgG H&L 800CW at 1/20000 dilution
Lanes 1 - 4:
Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution
Observed band size: 100 kDa
false
- NGS
Lab
Next Generation Sequencing - Human DGCR8 knockout HCT116 cell line (AB287366)
49 bp deletion (allele 1), 1 bp insertion and 13 bp deletion (allele 2) in exon 4, CCDS13773.1
Reactivity data
Product details
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
DGCR8 partners with Drosha to form the Microprocessor complex an important player in miRNA biogenesis. This complex identifies and accurately processes primary transcripts into precursor miRNAs a step essential for proper gene regulation. DGCR8's role is critical for maintaining the normal cellular function enabling the precise control of miRNA production which in turn modulates gene expression post-transcriptionally. Aberrations in its function affect the regulation of genes involved in cell cycle differentiation and development.
Pathways
DGCR8 holds a significant influence on the RNA interference (RNAi) pathway interacting closely with Drosha to regulate miRNA synthesis. This pathway is fundamental for the regulation of genetic networks involved in cellular development and homeostasis. In the context of the miRNA processing pathway DGCR8 links to proteins like Dicer which further processes precursor miRNAs in the cytoplasm. Proper function of these pathways ensures the balanced expression of mRNAn important for healthy cellular activities.
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
Complementary Products
![Western blot - Anti-DGCR8 antibody [EPR18757] (AB191875)](https://content.abcam.com/products/images/dgcr8-antibody-epr18757-ab191875--western-blot-img103531.jpg)
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