Human NONO (nmt55 / p54nrb) knockout HEK-293T cell line
- Advanced Validation
- What is this?
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(1 Publication)
- WB
Lab
Western blot - Human NONO (nmt55 / p54nrb) knockout HEK-293T cell line (AB266244)
Lanes 1-3 : Merged signal (red and green). Green - ab133574 observed at 63 kDa. Red - loading control, ab8245 observed at 37 kDa.
ab133574 Anti-nmt55 / p54nrb antibody [EPR5270] was shown to specifically react with nmt55 / p54nrb in wild-type HEK293T cells. Loss of signal was observed when knockout cell line ab266244 (knockout cell lysate ab257160) was used. Wild-type and nmt55 / p54nrb knockout samples were subjected to SDS-PAGE. ab133574 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 10000 dilution for 1 hour at room temperature before imaging.
All lanes:
Western blot - Anti-nmt55 / p54nrb antibody [EPR5270] (<a href='/en-us/products/primary-antibodies/nmt55-p54nrb-antibody-epr5270-ab133574'>ab133574</a>) at 1/1000 dilution
Lane 1:
Wild-type HEK293T cell lysate at 20 µg
Lane 2:
Western blot - Human NONO (nmt55 / p54nrb) knockout HEK-293T cell lysate (<a href='/en-us/products/cell-lysates/human-nono-nmt55-p54nrb-knockout-hek-293t-cell-lysate-ab257160'>ab257160</a>) at 20 µg
Lane 3:
MOLT-4 cell lysate at 20 µg
Secondary
Lanes 1 - 3:
Western blot - Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-irdye-800cw-preadsorbed-ab216773'>ab216773</a>)
Lanes 1 - 3:
Western blot - Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-mouse-igg-h-l-irdye-680rd-preadsorbed-ab216776'>ab216776</a>)
Predicted band size: 54 kDa
Observed band size: 63 kDa,37 kDa
false
- Sanger seq
Unknown
Sanger Sequencing - Human NONO (nmt55 / p54nrb) knockout HEK-293T cell line (AB266244)
Allele-2 : 17 bp deletion in exon 2.
- Sanger seq
Unknown
Sanger Sequencing - Human NONO (nmt55 / p54nrb) knockout HEK-293T cell line (AB266244)
Allele-1 : 25 bp deletion in exon2
Complementary Products
Primary Antibodies
AB133574
Anti-nmt55 / p54nrb antibody [EPR5270]
RabMAb|Recombinant|KO Validated
![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-nmt55 / p54nrb antibody [EPR5270] (AB133574)](https://content.abcam.com/products/images/nmt55-p54nrb-antibody-epr5270-ab133574--immunohistochemistry-formalin-pfa-fixed-paraffin-embedded-sections-img135545.jpg)
- 0
- 0
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
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
P54nrb is a multifunctional protein critical in nuclear processes. It forms part of a complex involved in RNA processing and nuclear retention of edited RNA. This complex also includes the proteins PSF and PSPC1 contributing to its role in transcription regulation and RNA stability. p54nrb impacts gene expression through its ability to bind to DNA and RNA influencing numerous regulatory pathways within the cell.
Pathways
P54nrb has essential roles in gene expression and RNA maturation pathways. It closely interacts with the RNA polymerase II machinery affecting transcription elongation. The protein has an established connection with the steroid receptor RNA activator (SRA) to modulate transcriptional responses. It is related to other DBHS proteins such as PSF which work together for coordinate regulation of gene expression and splice site selection.
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
Publications (1)
Recent publications for all applications. Explore the full list and refine your search
Journal of Cancer 12:1073-1084 PubMed33442405
2021
Applications
Unspecified application
Species
Unspecified reactive species
Product promise
Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.
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