HNRNPA1 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 insertion in exon 1 and 4 bp insertion in exon 1.
HNRPA1, Helix-destabilizing protein, Heterogeneous nuclear ribonucleoprotein A1, Heterogeneous nuclear ribonucleoprotein A1B protein, Heterogeneous nuclear ribonucleoprotein B2 protein, Heterogeneous nuclear ribonucleoprotein core protein A1, MGC102835, Nuclear ribonucleoprotein particle A1 protein, ROA1_HUMAN, Single strand DNA binding protein UP1, Single-strand RNA-binding protein, hnRNP A1, hnRNP core protein A1
HNRNPA1 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 insertion in exon 1 and 4 bp insertion in exon 1.
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.
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.
Heterogeneous nuclear ribonucleoprotein A1 (hnRNP A1) also known just as hnRNP A1 is a multifunctional protein that plays important roles in RNA processing mechanisms. It is a member of the hnRNP protein family involved in the packaging of nascent pre-mRNA. hnRNP A1 has a molecular mass of approximately 34-38 kDa and shows a widespread expression in various cells and tissues. This protein is recognized for its ability to bind to RNA molecules contributing to their proper processing splicing and transport within the cellular context.
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.
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.
HnRNP A1 is notably related to neurodegenerative diseases like Amyotrophic Lateral Sclerosis (ALS) and certain types of cancer. Abnormalities in its expression or mutations can lead to disrupted cellular processes contributing to disease pathology. In ALS dysregulation of hnRNP A1 affects neuronal RNA metabolism and is associated with the TDP-43 protein a marker of neurodegeneration. In cancer hnRNP A1 influences tumor development through its role in alternative splicing and gene expression modification linking it to oncogenic signaling pathways.
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Lanes 1-3: Merged signal (red and green). Green - Anti-hnRNP A1 antibody [EPR12768] ab177152 observed at 37 kDa. Red - loading control Anti-GAPDH antibody [6C5] - Loading Control ab8245 observed at 36 kDa.
Anti-hnRNP A1 antibody [EPR12768] 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 Human HNRNPA1 knockout HEK-293T cell lysate ab256942) was used. Wild-type and hnRNP A1 knockout samples were subjected to SDS-PAGE. Anti-hnRNP A1 antibody [EPR12768] ab177152 and Anti-GAPDH antibody [6C5] - Loading Control (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 (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (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] (Anti-hnRNP A1 antibody [EPR12768] ab177152) 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
All lanes: Western blot - Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) 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
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.
Allele-1: 1 bp insertion in exon 1
Allele-2: 4 bp insertion in exon 1.
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