RALA KO cell line available to order. KO validated by Western blot. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, Homozygous: 1 bp deletion in exon 2.
MGC48949, RAL, RALA_HUMAN, RAS like protein A, Ral A protein, Ras family small GTP binding protein RALA, Ras-related protein Ral-A, v ral simian leukemia viral oncogene homolog A, v ral simian leukemia viral oncogene homolog A (ras related)
RALA KO cell line available to order. KO validated by Western blot. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, Homozygous: 1 bp deletion in exon 2.
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 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.
RALA also known as Ras-related protein Ral-A or RalA is a small GTPase involved in a variety of cellular processes. It has a molecular mass of approximately 23 kDa. RALA switches between inactive GDP-bound and active GTP-bound states facilitating the transduction of signals within the cell. This protein is found in a range of tissues with expression in the brain lung and kidney being notable. The ability of RALA to shuttle between membrane compartments highlights its mechanical versatility in cellular signaling.
The interaction of RALA with effector proteins drives key processes like vesicle trafficking cytoskeletal dynamics and gene expression. RALA forms part of a signaling complex in its active state that mediates these functions. This protein is vital in the neurotransmitter release ensuring efficient communication between neurons. Additionally RALA impacts cell movement by influencing actin filament organization showing its role in cell motility.
The engagement of RALA in Ral signaling and the phosphatidylinositol 3-kinase (PI3K) pathway showcases its importance. In Ral signaling RALA interacts closely with proteins like RalBP1 influencing endocytosis and exocytosis activities. In the PI3K pathway RALA helps regulate cell proliferation and survival showing interactions with proteins such as AKT1. These pathways highlight the integration of RALA into broad signal transduction networks important for cellular homeostasis.
The altered regulation of RALA links it to cancer and neuronal disorders. In cancer the dysregulation of RALA can drive tumorigenesis often acting alongside proteins like KRAS in promoting oncogenic signaling. In neuronal disorders improper functioning of RALA has been associated with synaptic dysfunctions potentially affecting proteins like Synapsin I which are important to synaptic vesicle trafficking. Understanding RALA's role in these contexts helps in developing targeted therapeutic strategies.
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Anti-RALA antibody [EPR6468] ab126627 Anti-RALA antibody [EPR6468] was shown to specifically react with RALA in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab265092 (knockout cell lysate Human RALA knockout HeLa cell lysate ab258165) was used. Wild-type and RALA knockout samples were subjected to SDS-PAGE. Anti-RALA antibody [EPR6468] ab126627 and Anti-alpha Tubulin antibody [DM1A] - Loading Control (Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291) were incubated overnight at 4° 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-RALA antibody [EPR6468] (Anti-RALA antibody [EPR6468] ab126627) at 1/1000 dilution
Lane 1: Wild-type HeLa cell lysate at 20 µg
Lane 2: RALA knockout HeLa cell lysate at 20 µg
Lane 2: Western blot - Human RALA knockout HeLa cell line (ab265092)
Lane 3: MCF7 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 1/20000 dilution
Performed under reducing conditions.
Predicted band size: 24 kDa
Observed band size: 25 kDa
Homozygous: 1 bp deletion in exon 2.
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