Human DDX17 knockout HEK-293 cell line
- Advanced Validation
- What is this?
Be the first to review this product! Submit a review
|
(0 Publication)
- WB
Lab
Western blot - Human DDX17 knockout HEK-293 cell line (AB261721)
Lanes 1 - 3 : Merged signal (red and green). Green - ab71958 observed at 72 85 kDa. Red - loading control ab181602 observed at 37 kDa.
ab71958 was shown to recognize DDX17 in wild-type HEK-293 cells as signal was lost at the expected MW in DDX17 knockout cells. Additional cross-reactive bands were observed in the wild-type and knockout cells. Wild-type and DDX17 knockout samples were subjected to SDS-PAGE. ab71958 and ab181602 (Rabbit anti-GAPDH loading control) were incubated overnight at 4°C at 1/1000 dilution and 1/20000 dilution respectively. Blots were developed with Goat anti-Mouse IgG H&L (IRDye® 800CW) preabsorbed ab216772 and Goat anti-Rabbit IgG H&L (IRDye® 680RD) preabsorbed ab216777 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.
All lanes:
Western blot - Anti-DDX17 antibody [2248C2a] (<a href='/en-us/products/primary-antibodies/ddx17-antibody-2248c2a-ab71958'>ab71958</a>) at 1/1000 dilution
Lane 1:
Wild-type HEK-293 whole cell lysate at 20 µg
Lane 2:
DDX17 knockout HEK-293 whole cell lysate at 20 µg
Lane 2:
Western blot - Human DDX17 knockout HEK-293 cell line (ab261721)
Lane 3:
HeLa whole cell lysate at 20 µg
Predicted band size: 72 kDa
false
- WB
Lab
Western blot - Human DDX17 knockout HEK-293 cell line (AB261721)
Lanes 1 - 4 : Merged signal (red and green). Green - ab24601 observed at 72 kDa. Red - loading control ab8245 observed at 37 kDa.
ab24601 was shown to recognize DDX17 in wild-type HEK 293 cells as signal was lost at the expected MW in DDX17 knockout cells. Additional cross-reactive bands were observed in the wild-type and knockout cells. Wild-type and DDX17 knockout samples were subjected to SDS-PAGE. ab24601 and ab8245 (Mouse anti-GAPDH loading control) were incubated overnight at 4°C at 1 μg/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-DDX17 antibody (<a href='/en-us/products/primary-antibodies/ddx17-antibody-ab24601'>ab24601</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:
DDX17 knockout HEK-293 (Human epithelial cell line from embryonic kidney) whole cell lysate at 20 µg
Lane 2:
Western blot - Human DDX17 knockout HEK-293 cell line (ab261721)
Lane 3:
HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate at 20 µg
Predicted band size: 72 kDa
false
- WB
Lab
Western blot - Human DDX17 knockout HEK-293 cell line (AB261721)
Lanes 1 - 4 : Merged signal (red and green). Green - ab180190 observed at 72 kDa. Red - loading control ab8245 observed at 37 kDa.
ab180190 was shown to recognize DDX17 in wild-type HEK 293 cells as signal was lost at the expected MW in DDX17 knockout cells. Additional cross-reactive bands were observed in the wild-type and knockout cells. Wild-type and DDX17 knockout samples were subjected to SDS-PAGE. The membrane was blocked with 3% Milk. ab180190 and ab8245 (Mouse anti-GAPDH loading control) were incubated overnight at 4°C at 1/5000 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-DDX17 antibody [EPR13807(B)] (<a href='/en-us/products/primary-antibodies/ddx17-antibody-epr13807b-ab180190'>ab180190</a>) at 1/5000 dilution
Lane 1:
Wild-type HEK-293 (Human epithelial cell line from embryonic kidney) whole cell lysate at 20 µg
Lane 2:
DDX17 knockout HEK-293 (Human epithelial cell line from embryonic kidney) whole cell lysate at 20 µg
Lane 2:
Western blot - Human DDX17 knockout HEK-293 cell line (ab261721)
Lane 3:
HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate at 20 µg
Predicted band size: 72 kDa
false
- NGS
Supplier Data
Next Generation Sequencing - Human DDX17 knockout HEK-293 cell line (AB261721)
1 bp insertion after Gln215 of the WT protein
Complementary Products
Primary Antibodies
AB109012
Anti-SQSTM1 / p62 antibody [EPR4844] - Autophagosome Marker
KO Validated|RabMAb|Recombinant|Lab Essentials
![Western blot - Anti-SQSTM1 / p62 antibody [EPR4844] - Autophagosome Marker (AB109012)](https://content.abcam.com/products/images/sqstm1-p62-antibody-epr4844-autophagosome-marker-ab109012--western-blot-img100339.jpg)
- 5
- 15
Primary Antibodies
AB18207
Anti-beta III Tubulin antibody - Neuronal Marker
BOND RX™ Validated|Lab Essentials|KO Validated
- 5
- 67
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, ERS Genomics Limited and Sigma-Aldrich Co. LLC, and is developed with patented technology. For full details of the licenses and 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
This enzyme plays a significant role in RNA metabolism. DDX17 is part of the spliceosome complex which is essential for splicing pre-mRNA into mature mRNA. Through its helicase activity DDX17 ensures the proper remodeling of RNA structures which is necessary for accurate splicing. The protein also contributes to the regulation of gene expression and has been observed to influence transcription factors.
Pathways
RNA processing and maturation are key biological functions of DDX17. The protein integrates into the mRNA splicing pathway where it orchestrates proper RNA folding and splice site selection. Furthermore DDX17 interacts with other DEAD-box proteins such as DDX5 to assist in chromatin remodeling and transcription regulation facilitating efficient 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
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