Human HNRNPD knockout U-2 OS cell line
- Advanced Validation
- What is this?
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HNRNPD KO cell line available to order. KO validated by Next Generation Sequencing, Western blot. Free of charge wild type control available. Knockout achieved by CRISPR/Cas9 X = 5 bp deletion Frameshift: 100%. To order both knockout and wild-type control cells: select '2 x 1000000 Cells/vial'. To order only knockout cells: select '1000000 Cells/vial'.
View Alternative Names
ARE binding protein AUFI type A, AU-rich element RNA-binding protein 1, AUF, AUF1, AUF1A, HNRPD_HUMAN, Heterogeneous nuclear ribonucleoprotein D0, P37, hnRNP D, hnRNP D0
- WB
Lab
Western blot - Human HNRNPD knockout U-2 OS cell line (AB273860)
False colour image of Western blot : Anti-hnRNP D/AUF1 antibody [EPR24001-12] staining at 1/1000 dilution shown in green; loading control ab7291 (Mouse anti-Alpha Tubulin [DM1A]) staining at 1/20000 dilution shown in red. In Western blot ab259895 was shown to bind specifically to hnRNP D/AUF1. A band was observed at 40 kDa in wild-type U-2 OS cell lysates with no signal observed at this size in HNRNPD knockout cell line ab273860 (knockout cell lysate ab273814). To generate this image wild-type and HNRNPD knockout U-2 OS 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 °. 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 (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) at 1/20000 dilution.
All lanes:
Western blot - Anti-hnRNP D/AUF1 antibody [EPR24001-12] (<a href='/en-us/products/primary-antibodies/hnrnp-d-auf1-antibody-epr24001-12-ab259895'>ab259895</a>) at 1/1000 dilution
Lane 1:
Wild-type U-2 OS cell lysate at 20 µg
Lane 2:
HNRNPD knockout U-2 OS cell lysate at 20 µg
Lane 2:
Western blot - Human HNRNPD knockout U-2 OS cell line (ab273860)
Predicted band size: 38 kDa
Observed band size: 40 kDa
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- WB
Lab
Western blot - Human HNRNPD knockout U-2 OS cell line (AB273860)
Lanes 1 - 4 : Merged signal (red and green). Green - ab50692 observed at 40 kDa. Red - loading control ab7291 (Mouse anti-Alpha Tubulin [DM1A] observed at 55kDa.
ab50692 was shown to react with hnRNP D/AUF1 in wild-type U-2 OS cells in western blot with loss of signal observed in HNRNPD knockout cell line ab273860 (knockout cell lysate ab273814). Wild-type and HNRNPD knockout U-2 OS cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3% milk in TBS-T (0.1% Tween®) before incubation with ab50692 and ab7291 (Mouse anti-Alpha Tubulin [DM1A] overnight at 4°C at 1.25 μg/ml and a 1 in 20000 dilution respectively. Blots were incubated 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 in 20000 dilution for 1 hour at room temperature before imaging.
All lanes:
Anti-hnRNP D/AUF1 antibody (<a href='/en-us/products/unavailable/hnrnp-dauf1-antibody-ab50692'>ab50692</a>) at 1.25 µg/mL
Lane 1:
Wild-type U-2 OS (Human bone osteosarcoma epithelial cell line) whole cell lysate at 20 µg
Lane 2:
HNRNPD knockout U-2 OS (Human bone osteosarcoma epithelial cell line) whole cell lysate at 20 µg
Lane 2:
Western blot - Human HNRNPD knockout U-2 OS cell line (ab273860)
Lane 3:
Jurkat (Human T cell leukemia cell line from peripheral blood) whole cell lysate at 20 µg
Lane 4:
HEK-293 (Human epithelial cell line from embryonic kidney) whole cell lysate at 20 µg
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- NGS
Supplier Data
Next Generation Sequencing - Human HNRNPD knockout U-2 OS cell line (AB273860)
5 bp deletion after Pro83 of the WT protein
- NGS
Supplier Data
Next Generation Sequencing - Human HNRNPD knockout U-2 OS cell line (AB273860)
Knockout achieved by CRISPR/Cas9; X = 5 bp deletion; Frameshift : 100%
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 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
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
HnRNP D/AUF1 regulates the degradation of mRNA by binding to AU-rich elements (AREs) found in the 3' untranslated region of many genes. It is an integral part of RNA-protein complexes that are important for controlling the half-life of mRNAs. Besides mRNA decay hnRNP D/AUF1 participates in other processes like mRNA splicing and transport. Its interactions within the ribonucleoprotein complexes highlight its versatile role in RNA metabolism.
Pathways
HnRNP D/AUF1 operates within important biological processes such as the mRNA decay pathway and the stress response pathway. It interacts closely with other proteins involved in mRNA decay such as tristetraprolin and members of the poly-A binding protein family. hnRNP D/AUF1 influences the decay rate of different transcripts contributing to cellular responses to environmental stimuli and maintaining homeostasis.
Quality control
STR analysis
CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
Biosafety level
EU: 1 US: 1
Adherent/suspension
Adherent
Gender
Female
Target data
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Primary Antibodies
AB259895
Anti-hnRNP D/AUF1 antibody [EPR24001-12]
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Primary Antibodies
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Primary Antibodies
AB7291
Anti-alpha Tubulin antibody [DM1A] - Loading Control
BOND RX™ Validated
![Western blot - Anti-alpha Tubulin antibody [DM1A] - Loading Control (AB7291)](https://content.abcam.com/products/images/alpha-tubulin-antibody-dm1a-loading-control-ab7291--western-blot-img3844.jpg)
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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