ARF1 KO cell lysate available now. KO validated by Western blot. Free of charge wild type control included. Knockout achieved by using CRISPR/Cas9, 1 bp deletion in exon 2 and 1 bp insertion in exon 2.
ADP-ribosylation factor 1, ARF1_HUMAN
ARF1 KO cell lysate available now. KO validated by Western blot. Free of charge wild type control included. Knockout achieved by using CRISPR/Cas9, 1 bp deletion in exon 2 and 1 bp insertion in exon 2.
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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.
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ARF1 also known as ADP-ribosylation factor 1 or ARF1 protein is a small GTPase with a molecular mass of approximately 20 kDa. Commonly expressed in a variety of cell types ARF1 plays a central role in vesicle trafficking. As an activator ARF1 interacts with different GTPase-activating proteins such as ARFGAP1 and BIG1 ensuring the proper distribution of proteins and lipids in cells. ARF1's expression occurs across numerous cellular compartments including the Golgi apparatus aiding in the formation of transport vesicles.
ARF1 is essential in regulating membrane dynamics and vesicular traffic. It forms part of the COPI and clathrin-coated vesicle complexes where it recruits coat proteins to budding vesicles. This recruitment is fundamental for maintaining Golgi structure and function. Additionally ARF1 plays a role in cytokinesis by interacting with the septin cytoskeleton. The protein also influences actin cytoskeleton remodeling which is pivotal for cell shape changes and motility.
ARF1 is central to both the endocytic and secretory pathways. It collaborates with proteins like ARFS and 3F1 in modulating the trafficking of cargo between the endoplasmic reticulum and Golgi. Within the secretory pathway ARF1 interacts with SNARE proteins to facilitate vesicle docking and fusion. Its actions in pathways maintain cellular homeostasis and promote proper cellular response to various stimuli.
ARF1 has links to cancer and Alzheimer's disease. Overexpression of ARF1 correlates with tumor progression and metastasis impacting cell proliferation and survival mechanisms. Additionally its disruption associates with Alzheimer's where it may influence amyloid precursor protein processing alongside interactions with GAP proteins. Understanding ARF1's role in these conditions highlights its potential as a therapeutic target stressing the importance of studying its interactions with disease-related proteins.
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Lanes 1 - 4: Merged signal (red and green). Green - Anti-ARF1 antibody ab183576 observed at 18 kDa. Red - loading control, Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291 (Mouse anti-Alpha Tubulin [DM1A]) observed at 55kDa.
Anti-ARF1 antibody ab183576 was shown to react with ARF1 in wild-type HeLa cells in Western blot with loss of signal observed in ARF1 knockout cell line Human ARF1 knockout HeLa cell line ab264939 (ARF1 knockout cell lysate ab257353). Wild-type HeLa and ARF1 knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3 % milk in TBS-T (0.1 % Tween®) before incubation with Anti-ARF1 antibody ab183576 and Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291 (Mouse anti-Alpha Tubulin [DM1A]) overnight at 4 °C at a 1 in 1000 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 h at room temperature before imaging.
All lanes: Western blot - Anti-ARF1 antibody (Anti-ARF1 antibody ab183576) at 1/1000 dilution
Lane 1: Wild-type HeLa cell lysate at 20 µg
Lane 2: ARF1 knockout HeLa cell lysate at 20 µg
Lane 2: Western blot - Human ARF1 knockout HeLa cell line (Human ARF1 knockout HeLa cell line ab264939)
Lane 3: MDA-MB-231 cell lysate at 20 µg
Lane 4: PANC-1 cell lysate at 20 µg
Performed under reducing conditions.
Predicted band size: 21 kDa
Observed band size: 18 kDa
Lanes 1 - 2: Merged signal (red and green). Green - Anti-ARF1 antibody ab58578 observed at 18 kDa. Red - loading control Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291 (Mouse anti-Alpha Tubulin [DM1A]) observed at 55 kDa.
Anti-ARF1 antibody ab58578 was shown to react with ARF1 in wild-type HeLa cells in Western blot with loss of signal observed in ARF1 knockout cell line Human ARF1 knockout HeLa cell line ab264939 (ARF1 knockout cell lysate ab257353). Wild-type HeLa and ARF1 knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3 % milk in TBS-T (0.1 % Tween®) before incubation with Anti-ARF1 antibody ab58578 and Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291 (Mouse anti-Alpha Tubulin [DM1A]) overnight at 4 °C at 1 μg/ml and a 1 in 20000 dilution respectively. Blots were incubated with Donkey anti-Goat IgG H&L (IRDye® 800CW) preabsorbed (Donkey anti-Goat IgG H&L (IRDye® 800CW) preadsorbed ab216775) and Donkey anti-Mouse 680RD secondary antibodies at 1 in 20000 dilution for 1 h at room temperature before imaging.
All lanes: Western blot - Anti-ARF1 antibody (Anti-ARF1 antibody ab58578) at 1 µg/mL
Lane 1: Wild-type HeLa cell lysate at 20 µg
Lane 2: ARF1 knockout HeLa cell lysate at 20 µg
Lane 2: Western blot - Human ARF1 knockout HeLa cell line (Human ARF1 knockout HeLa cell line ab264939)
Performed under reducing conditions.
Predicted band size: 21 kDa
Observed band size: 18 kDa
Allele-1: 1 bp deletion in exon 2
Allele-2: 1 bp insertion in exon 2
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