HNRNPR KO cell line available to order. KO validated by Western blot. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, 1 bp deletion in exon 3 and 2 bp deletion in exon 3.
Images
Key facts
- Cell type
- HEK-293T
- Species or organism
- Human
- Tissue
- Kidney
- Form
- Liquid
- Knockout validation
- Sanger Sequencing, Western blot
- Mutation description
- Knockout achieved by using CRISPR/Cas9, 1 bp deletion in exon 3 and 2 bp deletion in exon 3
Alternative names
FLJ25714, HNRPR_HUMAN, Heterogeneous nuclear ribonucleoprotein R, hnRNP-R
Recommended products
HNRNPR KO cell line available to order. KO validated by Western blot. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, 1 bp deletion in exon 3 and 2 bp deletion in exon 3.
Key facts
- Cell type
- HEK-293T
- Species or organism
- Human
- Tissue
- Kidney
- Form
- Liquid
- Knockout validation
- Sanger Sequencing, Western blot
- Mutation description
- Knockout achieved by using CRISPR/Cas9, 1 bp deletion in exon 3 and 2 bp deletion in exon 3
- Concentration
Properties
- Gene name
- HNRNPR
- Gene editing type
- Knockout
- Gene editing method
- CRISPR technology
- Knockout validation
- Sanger Sequencing, Western blot
Quality control
- STR analysis
- CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
- Biosafety level
- EU: 2 US: 2
- Adherent/suspension
- Adherent
- Gender
- Female
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.
Storage
- Shipped at conditions
- Dry Ice
- Appropriate short-term storage conditions
- -196°C
- Appropriate long-term storage conditions
- -196°C
Notes
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.
Supplementary info
- This supplementary information is collated from multiple sources and compiled automatically.
- Activity summary
Heterogeneous nuclear ribonucleoprotein R commonly known as hnRNP R is a multifunctional protein involved in mRNA processing. It is approximately 72 kDa in size and found mostly in the nucleus. hnRNP R exhibits a strong expression in neural tissues and various other cell types. As a member of the hnRNP family it plays an important role in RNA-binding activities and influences the splicing process export and stability of mRNA.
- Biological function summary
HnRNP R contributes to several cellular tasks related to gene expression regulation. This protein interacts with other hnRNPs to form core components of ribonucleoprotein complexes. It participates in modulating RNA transport from the nucleus to the cytoplasm and facilitates mRNA translation. Additionally hnRNP R associates with other proteins and RNA molecules strengthening the cellular response to environmental cues.
- Pathways
HnRNP R acts decisively in several gene expression and RNA processing pathways. It is involved in the mRNA splicing pathway where it interacts with splicing machinery to process precursor mRNAs into mature mRNAs. hnRNP R also influences the Wnt signaling pathway by regulating the stability and localization of mRNAs involved in this pathway. Moreover it interacts with proteins like hnRNP A1 which further aids its functional capabilities in these pathways.
- Associated diseases and disorders
HnRNP R associates with neurodegenerative diseases like amyotrophic lateral sclerosis (ALS) and spinal muscular atrophy (SMA). Its interaction with other hnRNPs like TDP-43 suggests a link to the RNA-binding protein misregulation observed in these disorders. Research shows that abnormalities in hnRNP R function can lead to widespread disruption of mRNA processing which contributes to the pathogenesis of neurodegenerative diseases.
Product promise
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3 product images
Western blot - Human HNRNPR knockout HEK-293T cell line (ab266723)
False colour image of Western blot: Anti-hnRNP R antibody staining at 1/500 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (Anti-GAPDH antibody [6C5] - Loading Control ab8245) loading control staining at 1/20000 dilution, shown in red. In Western blot, Anti-hnRNP R antibody ab151251 was shown to bind specifically to hnRNP R. A band was observed at 78 kDa in wild-type HEK-293T cell lysates with no signal observed at this size in HNRNPR knockout cell line ab266723 (knockout cell lysate Human HNRNPR knockout HEK-293T cell lysate ab257992). To generate this image, wild-type and HNRNPR knockout HEK-293T 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 °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 (IRDye® 800CW) preabsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776) at 1/20000 dilution.
All lanes: Western blot - Anti-hnRNP R antibody (Anti-hnRNP R antibody ab151251) at 1/500 dilution
Lane 1: Wild-type HEK-293T cell lysate at 20 µg
Lane 2: HNRNPR knockout HEK-293T cell lysate at 20 µg
Lane 2: Western blot - Human HNRNPR knockout HEK-293T cell line (ab266723)
Lane 3: MOLT-4 cell lysate at 20 µg
Lane 4: Caco-2 cell lysate at 20 µg
Performed under reducing conditions.
Predicted band size: 71 kDa
Observed band size: 78 kDa
Sanger Sequencing - Human HNRNPR knockout HEK-293T cell line (ab266723)
Allele-1: 2 bp deletion in exon 3
Sanger Sequencing - Human HNRNPR knockout HEK-293T cell line (ab266723)
Allele-2: 1 bp deletion in exon 3.
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