Human DDX17 knockout HEK-293 cell line (ab261721)
Overview
-
Product name
Human DDX17 knockout HEK-293 cell line -
Parental Cell Line
HEK-293 -
Organism
Human -
Mutation description
Knockout achieved by CRISPR/Cas9; X = 1 bp insertion -
Passage number
<20 -
Knockout validation
Next Generation Sequencing (NGS), Western Blot (WB) -
Tested applications
Suitable for: WBmore details -
Biosafety level
2 -
General notes
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.
Cryopreservation cell medium: Cell Freezing Medium-DMSO Serum free media, contains 8.7% DMSO in MEM supplemented with methyl cellulose.
Culture medium: DMEM (High Glucose) + 10% FBS
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. Based on cell count, seed cells in an appropriate cell culture 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.
4. Incubate the culture at 37°C incubator with 5% CO2. 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.
- A partial media change 24 hours prior to subculture may be helpful to encourage growth, if required.
- Cells should be passaged when they have achieved 80-90% confluence.
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.
We will provide viable cells that proliferate on revival.
Properties
-
Number of cells
1 x 106 cells/vial, 1 mL -
Adherent /Suspension
Adherent -
Tissue
Kidney -
Cell type
epithelial -
Gender
Female -
Mycoplasma free
Yes -
Storage instructions
Shipped on Dry Ice. Store in liquid nitrogen. -
Storage buffer
Constituents: 8.7% Dimethylsulfoxide, 2% Cellulose, methyl ether -
Research areas
Target
-
Function
RNA-dependent ATPase activity. -
Tissue specificity
Ubiquitous. -
Sequence similarities
Belongs to the DEAD box helicase family. DDX5/DBP2 subfamily.
Contains 1 helicase ATP-binding domain.
Contains 1 helicase C-terminal domain. -
Post-translational
modificationsPhosphorylated upon DNA damage, probably by ATM or ATR. -
Cellular localization
Nucleus. - Information by UniProt
Associated products
-
KO cell lysates
-
Related Products
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab261721 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
| Application | Abreviews | Notes |
|---|---|---|
| WB |
Use at an assay dependent concentration. Predicted molecular weight: 72 kDa.
|
| Notes |
|---|
|
WB
Use at an assay dependent concentration. Predicted molecular weight: 72 kDa. |
Images
-
Western blot - Human DDX17 knockout HEK-293 cell line (ab261721)All lanes : Anti-DDX17 antibody [2248C2a] (ab71958) at 1/1000 dilution
Lane 1 : Wild-type HEK-293 whole cell lysate
Lane 2 : DDX17 knockout HEK-293 whole cell lysate
Lane 3 : HeLa whole cell lysate
Lysates/proteins at 20 µg per lane.
Predicted band size: 72 kDaLanes 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 cell line ab261721 (knockout cell lysate ab261660). Additional cross-reactive bands were observed in the wild-type and knockout samples. 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.
-
Western blot - Human DDX17 knockout HEK-293 cell line (ab261721)All lanes : Anti-DDX17 antibody (ab24601) at 1 µg/ml
Lane 1 : Wild-type HEK-293 (Human epithelial cell line from embryonic kidney) whole cell lysate
Lane 2 : DDX17 knockout HEK-293 (Human epithelial cell line from embryonic kidney) whole cell lysate
Lane 3 : HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 72 kDaLanes 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 cell line ab261721 (knockout cell lysate ab261660). Additional cross-reactive bands were observed in the wild-type and knockout cell lysate. 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.
-
Western blot - Human DDX17 knockout HEK-293 cell line (ab261721)All lanes : Anti-DDX17 antibody [EPR13807(B)] (ab180190) at 1/5000 dilution
Lane 1 : Wild-type HEK-293 (Human epithelial cell line from embryonic kidney) whole cell lysate
Lane 2 : DDX17 knockout HEK-293 (Human epithelial cell line from embryonic kidney) whole cell lysate
Lane 3 : HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 72 kDaLanes 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 cell line ab261721 (knockout cell lysate ab261660). Additional cross-reactive bands were observed in the wild-type and knockout cell lysate. 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.
Protocols
Datasheets and documents
-
SDS download
-
Datasheet download
References (0)
ab261721 has not yet been referenced specifically in any publications.