ACTA2 KO cell lysate available now. KO validated by Immunocytochemistry, Next Generation Sequencing, Western blot. Free of charge wild type control included.
AAT6, ACTA2, ACTA_HUMAN, ACTSA, ACTVS, Actin alpha 2 smooth muscle aorta, Actin aortic smooth muscle, Alpha 2 actin, Alpha cardiac actin, Alpha-actin-2, Cell growth-inhibiting gene 46 protein, GIG46, Growth inhibiting gene 46, MYMY5, a-actin, alpha sma
ACTA2 KO cell lysate available now. KO validated by Immunocytochemistry, Next Generation Sequencing, Western blot. Free of charge wild type control included.
Knockout cell lysate achieved by CRISPR/Cas9.
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Lysate preparation: Our lysates are made using RIPA buffer to which we add a protease inhibitor cocktail and phosphatase inhibitor cocktail (ratio: 300:100:10). This means that the protein of interest is denatured. If you require a native form of the protein please use the live cell version. Please refer to our lysis protocol for further details on how our lysates are prepared.
User storage instructions: Lyophilizate may be stored at 4°C. After reconstitution, store at -20°C for short-term storage or -80°C for long-term storage.
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.
Alpha smooth muscle actin (α-SMA) also known as ACTA2 is an actin isoform with a specific role in the contractile function of smooth muscle cells. The molecular weight of α-SMA is approximately 42 kDa. This protein is expressed widely in vascular smooth muscle cells the peritoneal lining and myofibroblasts. It often serves as a marker for these cell types. The expression of α-SMA is critical for the mechanical attributes of cells contributing to the rigidity and contractility of tissues where it is present.
Alpha smooth muscle actin aids in maintaining the structural integrity of tissues by forming part of the actin cytoskeleton an important element in cellular support. It exists in high concentration in stress fibers contributing to cellular movements and shape maintenance. These actions are essential in various dynamic cellular processes such as cell migration and adhesion. Alpha smooth muscle actin does not typically form complexes but it associates with other components of the actin cytoskeleton to ensure cell stability and function.
Alpha smooth muscle actin functions significantly within the TGF-beta signaling pathway which influences cell proliferation differentiation and apoptosis. It interacts closely with proteins such as myosin to facilitate cellular contractility and motility. Additionally α-SMA plays a part in the RhoA/Rho kinase (ROCK) pathway connecting with regulators of actin filament organization. These pathways are essential for modulation of smooth muscle contraction and actin filament assembly contributing to vascular development and wound healing.
Alpha smooth muscle actin is importantly involved in fibrotic diseases and vascular diseases like arteriosclerosis. Increased expression of α-SMA is often observed in fibrotic tissues contributing to pathogenesis due to excessive extracellular matrix deposition. In the context of vascular disorders overexpression of α-SMA can lead to abnormal vascular remodeling often seen in diseases like hypertension. Interactions with proteins such as fibronectin and collagens facilitate these pathological changes highlighting the importance of α-SMA in disease development.
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Lane 1: Wild-type HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate
Lane 2: ACTA2 knockout HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate
Lanes 1 - 2: Merged signal (red and green). Green - Anti-alpha smooth muscle Actin antibody [EPR5368] ab124964 observed at 42 kDa. Red - loading control, Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291 (Mouse anti-Alpha Tubulin [DM1A]) observed at 55kDa.
Anti-alpha smooth muscle Actin antibody [EPR5368] ab124964 was shown to react with alpha smooth muscle Actin in wild-type HeLa cells in western blot Loss of signal was observed when ACTA2 knockout cell line Human ACTA2 knockout HeLa cell line ab264014 (knockout cell lysate ab264499) was used. Wild-type HeLa and ACTA2 knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3% milk in TBS-T (0.1% Tween®) before incubation with Anti-alpha smooth muscle Actin antibody [EPR5368] ab124964 and Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291 (Mouse anti-Alpha Tubulin [DM1A]) overnight at 4°C at a 1 in 10000 dilution and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (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-alpha smooth muscle Actin antibody [EPR5368] (Anti-alpha smooth muscle Actin antibody [EPR5368] ab124964) at 1/10000 dilution
Lane 1: Wild-type HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate at 20 µg
Lane 2: ACTA2 knockout HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate at 20 µg
Lane 2: Western blot - Human ACTA2 knockout HeLa cell line (Human ACTA2 knockout HeLa cell line ab264014)
Performed under reducing conditions.
Predicted band size: 42 kDa
Observed band size: 42 kDa
Lane 1: Wild-type HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate
Lane 2: ACTA2 knockout HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate
HRP Anti-alpha smooth muscle Actin antibody [1A4] ab203696 was shown to react with alpha smooth muscle Actin (HRP) in wild-type HeLa cells in western blot. Loss of signal was observed when ACTA2 knockout cell line Human ACTA2 knockout HeLa cell line ab264014 (knockout cell lysate ab264499) was used. Membranes were blocked in 3% milk in TBS-T (0.1% Tween®) before incubation with HRP Anti-alpha smooth muscle Actin antibody [1A4] ab203696 overnight at 4°C at a 1 in 5000 dilution and Alexa Fluor® 680 Anti-GAPDH antibody [mAbcam 9484] - Loading Control ab184095 (Mouse Anti-GAPDH antibody [mAbcam 9484] - Alexa Fluor® 680) at a 1 in 1000 dilution. Blots were developed with Optiblot ECL reagent (Anti-PKC delta (phospho S299) antibody [EPNCI119] ab133456) and imaged.
All lanes: Western blot - HRP Anti-alpha smooth muscle Actin antibody [1A4] (HRP Anti-alpha smooth muscle Actin antibody [1A4] ab203696) at 1/5000 dilution
Lane 1: Wild-type HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate at 20 µg
Lane 2: ACTA2 knockout HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate at 20 µg
Performed under reducing conditions.
Predicted band size: 42 kDa
Observed band size: 42 kDa
Exposure time: 20s
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