Human HNRNPA2B1 knockout HEK-293T cell line
- Advanced Validation
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Supplier Data
Immunocytochemistry/ Immunofluorescence - Human HNRNPA2B1 knockout HEK-293T cell line (AB266404)
- WB
Lab
Western blot - Human HNRNPA2B1 knockout HEK-293T cell line (AB266404)
Blocking and diluting buffer and concentration : Intercept® (TBS) Blocking Buffer diluted with an equal volume of 0.1% TBS
Lanes 1- 4 : Merged signal (red and green). Green - ab259894 observed at 36/38kDa. Red - loading control ab7291 (Mouse monoclonal [DM1A] to alpha Tubulin) observed at 50 kDa.
Lanes 1-2 : ab259894 Anti-hnRNP A2B1 antibody was shown to react with hnRNP A2B1 in HEK-293T cells in Western blot. Loss of signal was observed when hnRNP A2B1 knockout cell line ab266404 (knockout cell lysate ab257224) was used. Wild-type and hnRNP A2B1 knockout samples were subjected to SDS-PAGE.
ab259894 and Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) were incubated at 4° overnight at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat Anti-Rabbit IgG H&L (IRDye® 800CW) (ab216773) and Goat Anti-Mouse IgG H&L (IRDye® 680RD) (ab216776) secondary antibodies at 1 in 10000 dilution for 1 hour at room temperature before imaging.
All lanes:
Western blot - Anti-hnRNP A2B1 antibody [EPR24002-81] (<a href='/en-us/products/primary-antibodies/hnrnp-a2b1-antibody-epr24002-81-ab259894'>ab259894</a>) at 1/1000 dilution
Lane 1:
Wild-type HEK-293T (human embryonic kidney epithelial cell), whole cell lysate at 20 µg
Lane 2:
hnRNP A2B1 knockout HEK-293T (human embryonic kidney epithelial cell), whole cell lysate at 20 µg
Lane 2:
Western blot - Human HNRNPA2B1 knockout HEK-293T cell line (ab266404)
Lane 3:
HeLa (human cervix adenocarcinoma epithelial cell), whole cell lysate at 20 µg
Lane 4:
A431 (human epidermoid carcinoma epithelial cell), whole cell lysate at 20 µg
Lane 5:
K562 (human chronic myelogenous leukemia lymphoblast), whole cell lysate at 20 µg
Secondary
All lanes:
Goat Anti-Rabbit IgG H&L (IRDye® 800CW) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-irdye-800cw-preadsorbed-ab216773'>ab216773</a>) and Goat Anti-Mouse IgG H&L (IRDye® 680RD) (<a href='/en-us/products/secondary-antibodies/goat-mouse-igg-h-l-irdye-680rd-preadsorbed-ab216776'>ab216776</a>) at 1/10000 dilution
Predicted band size: 37 kDa
Observed band size: 36 kDa,38 kDa
false
- WB
Lab
Western blot - Human HNRNPA2B1 knockout HEK-293T cell line (AB266404)
Lanes 1-3 : Merged signal (red and green). Green - ab31645 observed at 37 kDa. Red - loading control ab7291 observed at 50 kDa.
ab31645 Anti-hnRNP A2B1 antibody was shown to specifically react with hnRNP A2B1 in wild-type HEK293T cells. Loss of signal was observed when knockout cell line ab266404 (knockout cell lysate ab257224) was used. Wild-type and hnRNP A2B1 knockout samples were subjected to SDS-PAGE. ab31645 and Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) were incubated overnight at 4° at 1 in 500 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-hnRNP A2B1 antibody (<a href='/en-us/products/primary-antibodies/hnrnp-a2b1-antibody-ab31645'>ab31645</a>) at 1/500 dilution
Lane 1:
Wild-type HEK293T cell lysate
Lane 2:
HNRNPA2B1 knockout HEK293T cell lysate
Lane 2:
Western blot - Human HNRNPA2B1 knockout HEK-293T cell line (ab266404)
Lane 3:
A549 cell lysate at 20 µg
Predicted band size: 37 kDa
Observed band size: 37 kDa
false
- Cell Culture
Unknown
Cell Culture - Human HNRNPA2B1 knockout HEK-293T cell line (AB266404)
Representative images of HNRNPA2B1 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.
- Sanger seq
Unknown
Sanger Sequencing - Human HNRNPA2B1 knockout HEK-293T cell line (AB266404)
Homozygous : 2 bp deletion in exon 3
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Cell Lines & Lysates
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Human CAV1 (Caveolin-1) knockout HeLa cell line
Advanced Validation
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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
Zygosity
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 A2B1 contributes to alternative splicing RNA stability and translational regulation. This protein typically associates with complex assemblies that manage RNA processing and transport mechanisms. It interacts with other proteins within the heterogeneous nuclear ribonucleoprotein family collaborating in the regulation of mRNA cycling between the nucleus and cytoplasm affecting gene expression patterns.
Pathways
HnRNP A2B1 exhibits critical involvement in the mRNA splicing pathway and RNA transport pathways. It frequently interacts with proteins such as hnRNP A1 and hnRNP C where they collectively influence pre-mRNA processing and splicing dictating proper RNA maturation and cell function. These pathways are important for maintaining cellular homeostasis and responding to cellular signals.
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