Mouse Monoclonal beta Actin antibody. Suitable for Flow Cyt (Intra), WB, ICC/IF and reacts with Human, Mouse samples. Cited in 28 publications.
pH: 7.4
Preservative: 0.02% Sodium azide
Constituents: 99% PBS
Flow Cyt (Intra) | WB | ICC/IF | |
---|---|---|---|
Human | Tested | Expected | Tested |
Mouse | Expected | Tested | Expected |
Species | Dilution info | Notes |
---|---|---|
Species Human | Dilution info 0.10000-1.00000 µg for 106 Cells | Notes Mouse IgM [B11/7] - Isotype control ab91545 - Mouse monoclonal IgM, is suitable for use as an isotype control with this antibody. |
Species | Dilution info | Notes |
---|---|---|
Species Mouse | Dilution info Use at an assay dependent concentration. | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Mouse | Dilution info 1/500 | Notes Abcam recommends blocking with 3% Milk |
Species | Dilution info | Notes |
---|---|---|
Species Human | Dilution info Use at an assay dependent concentration. | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Human | Dilution info 5 µg/mL | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Mouse | Dilution info Use at an assay dependent concentration. | Notes - |
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Actin is a highly conserved protein that polymerizes to produce filaments that form cross-linked networks in the cytoplasm of cells (PubMed:25255767, PubMed:29581253). Actin exists in both monomeric (G-actin) and polymeric (F-actin) forms, both forms playing key functions, such as cell motility and contraction (PubMed:29581253). In addition to their role in the cytoplasmic cytoskeleton, G- and F-actin also localize in the nucleus, and regulate gene transcription and motility and repair of damaged DNA (PubMed:29925947). Part of the ACTR1A/ACTB filament around which the dynactin complex is built. The dynactin multiprotein complex activates the molecular motor dynein for ultra-processive transport along microtubules (By similarity).
Beta-actin, ACTB
Mouse Monoclonal beta Actin antibody. Suitable for Flow Cyt (Intra), WB, ICC/IF and reacts with Human, Mouse samples. Cited in 28 publications.
pH: 7.4
Preservative: 0.02% Sodium azide
Constituents: 99% PBS
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This antibody clone is manufactured by Abcam. If you require a custom buffer formulation or conjugation for your experiments, please contact orders@abcam.com
F-actin also known as filamentous actin is an essential structural protein found within the cytoskeleton of eukaryotic cells. Its alternate names include actin filaments or microfilaments. The protein consists of polymerized monomers of G-actin each with a molecular weight of roughly 42 kDa. F-actin is expressed abundantly in muscle cells and non-muscle cells alike providing structural support and facilitating cellular movements. Actin staining is a common method used in labs to visualize these dynamic structures often employing phalloidin staining a toxin that stabilizes actin filaments conjugated with fluorescent labels such as Phalloidin 594 Phalloidin 647 or Phalloidin 488 for imaging purposes.
The actin cytoskeleton plays integral roles in maintaining cell shape providing mechanical resistance against deformation and driving important cellular processes such as endocytosis cell division and motility. F-actin forms part of numerous protein complexes interacting with other proteins like myosin to facilitate muscle contraction and cellular transport. Within cells F-actin is dynamic readily polymerizing and depolymerizing in response to cellular signaling making it essential for cytoskeletal remodeling and cellular adaptability.
F-actin is central to various signaling cascades underlying processes like cell signaling and intracellular transport. Notably it participates in the Rho family GTPase pathway affecting cell cytoskeleton organization and motility. It also interacts with proteins like cofilin and profilin which regulate actin polymerization and treadmilling dynamics respectively. These interactions highlight F-actin's involvement in complex cellular pathway regulation processes essential for maintaining cellular homeostasis and adaptability.
Abnormal regulation or mutations in actin-related proteins can lead to conditions such as cancer and cardiomyopathies. For example during metastasis cancer cells exploit the dynamic nature of F-actin for enhanced migratory capacity. In cardiac muscle cells actin interacts with other proteins like tropomyosin and mutations in these genes can disrupt normal heart function leading to cardiomyopathies. As such F-actin not only represents a critical component of the cellular structure but also serves as a pivotal target for understanding disease mechanisms and potential therapeutic intervention points.
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This species and application combination has not been tested, but we predict it will work based on strong homology. However, this combination is not covered by our product promise.
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Overlay histogram showing HeLa cells stained with ab130935 (red line). The cells were fixed with 4% paraformaldehyde (10 min) and then permeabilized with 0.1% PBS-Triton X-100 for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab130935, 1μg/1x106 cells) for 30 min at 22°C. The secondary antibody used was a goat Anti-mouse DyLight® 488 (IgM; mu chain, Goat Anti-Mouse IgM mu chain (DyLight® 488) ab97007) at 1/2000 dilution for 30 min at 22°C. Isotype control antibody (black line) was mouse IgM [ICIGM] (Mouse IgM [B11/7] - Isotype control ab91545, 1μg/1x106 cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter. This antibody gave a positive signal in HeLa cells fixed with 80% methanol (5 min)/permeabilized with 0.1% PBS-Triton X-100 for 20 min used under the same conditions.
F-actin Western blot staining using mouse Anti-F-actin antibody
All lanes: Western blot - Anti-F-actin antibody [4E3.adl] - Loading Control (ab130935) at 1/500 dilution
Lane 1: Human skeletal muscle tissue lysate - total protein (ab29330) at 20 µg
Lane 2: Skeletal Muscle (Mouse) Tissue Lysate at 20 µg
Lane 3: Heart (Mouse) Tissue Lysate at 20 µg
Lane 4: HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate at 20 µg
Lane 5: C2C12 (Mouse myoblast cell line) Whole Cell Lysate at 20 µg
All lanes: Peroxidase- conjugated AffiniPure Goat Anti-mouse IgM (ab98112) at 1/3000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 41 kDa
Observed band size: 42 kDa, 60 kDa, 70 kDa
Exposure time: 10s
ab130935 stained HeLa cells. The cells were 100% methanol fixed (5 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody ab130935 at 5μg/ml overnight at +4°C. The secondary antibody (green) was a goat anti-mouse DyLight® 488 (Goat Anti-Mouse IgG H&L (DyLight® 488) preadsorbed ab96879) IgG (H+L) used at a 1/250 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43μM.
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