Human HNRNPA1 knockout HEK-293T cell line
- Advanced Validation
- What is this?
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(0 Publication)
- WB
Lab
Western blot - Human HNRNPA1 knockout HEK-293T cell line (AB266193)
Lanes 1-3 : Merged signal (red and green). Green - ab177152 observed at 37 kDa. Red - loading control ab8245 observed at 36 kDa.
ab177152 Anti-hnRNP A1 antibody [EPR12768] was shown to specifically react with hnRNP A1 in wild-type HEK293T cells. Loss of signal was observed when knockout cell line ab266193 (knockout cell lysate ab256942) was used. Wild-type and hnRNP A1 knockout samples were subjected to SDS-PAGE. ab177152 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 20000 dilution for 1 hour at room temperature before imaging.
All lanes:
Western blot - Anti-hnRNP A1 antibody [EPR12768] (<a href='/en-us/products/primary-antibodies/hnrnp-a1-antibody-epr12768-ab177152'>ab177152</a>) at 1/1000 dilution
Lane 1:
Wild-type HEK293T cell lysate at 20 µg
Lane 2:
HNRNPA1 knockout HEK293T cell lysate at 20 µg
Lane 2:
Western blot - Human HNRNPA1 knockout HEK-293T cell line (ab266193)
Lane 3:
HeLa cell lysate at 20 µg
Secondary
All lanes:
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>) at 1/10000 dilution
Predicted band size: 23 kDa,25 kDa,33 kDa,38 kDa,53 kDa,72 kDa
Observed band size: 25 kDa,27 kDa,37 kDa,38 kDa,58 kDa,72 kDa
false
- Sanger seq
Unknown
Sanger Sequencing - Human HNRNPA1 knockout HEK-293T cell line (AB266193)
Allele-2 : 4 bp insertion in exon 1.
- Sanger seq
Unknown
Sanger Sequencing - Human HNRNPA1 knockout HEK-293T cell line (AB266193)
Allele-1 : 1 bp insertion in exon 1
- Cell Culture
Unknown
Cell Culture - Human HNRNPA1 knockout HEK-293T cell line (AB266193)
Representative images of HNRNPA1 knockout HEK293T cells, low and high confluency examples (top left and right respectively) and wild-type HEK293T cells, low and high confluency (bottom left and right respectively) showing typical adherent, epithelial-like morphology. Images were captured at 10X magnification using a EVOS XL Core microscope.
Complementary Products
Cell Lines & Lysates
AB255371
Human CAV1 (Caveolin-1) knockout HeLa cell line
Advanced Validation
- 0
- 0
![Immunocytochemistry/ Immunofluorescence - Anti-hnRNP A1 antibody [EPR12768] (AB177152)](https://content.abcam.com/products/images/hnrnp-a1-antibody-epr12768-ab177152--immunocytochemistry-immunofluorescence-img1413.gif)
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
HnRNP A1 is involved in RNA binding and splicing contributing to the formation of spliceosomes the complexes responsible for pre-mRNA splicing. The protein aids in alternative splicing influencing mRNA diversity and stability. hnRNP A1 interacts with other proteins in the hnRNP family remodeling ribonucleoprotein complexes and affecting their functions. These interactions ensure proper mRNA maturation impacting gene expression regulation within the cell.
Pathways
HnRNP A1 is integral to critical cellular mechanisms such as the regulation of mRNA transport and splicing. It influences the DNA damage response pathway by modulating gene expression required for cellular repair and stability. hnRNP A1 also cooperates with other RNA binding proteins like the splicing factors SR proteins to orchestrate the splicing machinery. These interactions are pivotal in maintaining cellular homeostasis and response to stress.
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
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