Human DCP1A knockout HEK-293 cell line
- Advanced Validation
- What is this?
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DCP1A KO cell line available to order. KO validated by Next Generation Sequencing, Western blot. Free of charge wild type control provided. Knockout achieved by CRISPR/Cas9 X = 7 bp deletion Frameshift = 100%.
View Alternative Names
DCP1 decapping enzyme homolog A, DCP1A_HUMAN, Decapping enzyme hDcp1a, Decapping mRNA 1A, HSA275986, Nbla00360, Putative protein product of Nbla00360, SMAD4IP1, SMIF, Smad4-interacting transcriptional co-activator, Transcription factor SMIF, mRNA-decapping enzyme 1A
- WB
Lab
Western blot - Human DCP1A knockout HEK-293 cell line (AB261857)
Lanes 1 - 3 : Merged signal (red and green). Green - ab47811 observed at 75 kDa. Red - loading control ab8245 observed at 37 kDa.
ab47811 was shown to recognize DCP1A in wild-type HEK-293 cells as signal was lost at the expected MW in DCP1A knockout cell line ab261857 (knockout cell lysate ab261665). Additional cross-reactive bands were observed in the wild-type and knockout samples. Wild-type and DCP1A knockout samples were subjected to SDS-PAGE. ab47811 and ab8245 (Mouse anti GAPDH loading control) were incubated overnight at 4° at 1 ug/ml and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.
All lanes:
Western blot - Anti-Dcp1a antibody (<a href='/en-us/products/primary-antibodies/dcp1a-antibody-ab47811'>ab47811</a>) at 1 µg/mL
Lane 1:
Wild-type HEK-293 (Human epithelial cell line from embryonic kidney) whole cell lysate at 20 µg
Lane 2:
DCP1A knockout HEK-293 (Human epithelial cell line from embryonic kidney) whole cell lysate at 20 µg
Lane 2:
Western blot - Human DCP1A knockout HEK-293 cell line (ab261857)
Lane 3:
HepG2 (Human liver hepatocellular carcinoma cell line) whole cell lysate at 20 µg
Predicted band size: 63 kDa
Observed band size: 75 kDa
false
- WB
Lab
Western blot - Human DCP1A knockout HEK-293 cell line (AB261857)
Lanes 1 - 3 : Merged signal (red and green). Green - ab183709 observed at 75 kDa. Red - loading control ab8245 observed at 37 kDa.
ab183709 was shown to specifically react with DCP1A in wild-type HEK-293 cells as signal was lost in DCP1A knockout cell line ab261857 (knockout cell lysate ab261665). Wild-type and DCP1A knockout samples were subjected to SDS-PAGE. ab183709 and ab8245 (Mouse anti GAPDH loading control) were incubated overnight at 4° at 1/1000 dilution and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.
All lanes:
Western blot - Anti-Dcp1a antibody [EPR13822] (<a href='/en-us/products/primary-antibodies/dcp1a-antibody-epr13822-ab183709'>ab183709</a>) at 1/1000 dilution
Lane 1:
Wild-type HEK-293 (Human epithelial cell line from embryonic kidney) whole cell lysate at 20 µg
Lane 2:
DCP1A knockout HEK-293 (Human epithelial cell line from embryonic kidney) whole cell lysate at 20 µg
Lane 2:
Western blot - Human DCP1A knockout HEK-293 cell line (ab261857)
Lane 3:
HepG2 (Human liver hepatocellular carcinoma cell line) whole cell lysate at 20 µg
Predicted band size: 63 kDa
Observed band size: 75 kDa
false
- NGS
Lab
Next Generation Sequencing - Human DCP1A knockout HEK-293 cell line (AB261857)
X = 7 bp deletion
- NGS
Supplier Data
Next Generation Sequencing - Human DCP1A knockout HEK-293 cell line (AB261857)
7 bp deletion after Lys53 of the WT protein
Reactivity data
Product details
Recommended control: Human wild-type HEK-293 cell line (ab259776). 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.
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.
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
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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