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F-actin Staining Kit - Blue Fluorescence | Cytopainter (ab112124) fluorescence imaging kits are a set of fluorescence imaging tools for labeling sub-cellular organelles such as lysosomes, mitochondria, and actin filaments.


Images

Fluorescence Microscopy - F-actin Staining Kit - Blue Fluorescence - Cytopainter (AB112124), expandable thumbnail
  • Fluorescence Microscopy - F-actin Staining Kit - Blue Fluorescence - Cytopainter (AB112124), expandable thumbnail

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Key facts

Sample types
Tissue, Suspension cells, Adherent cells
Assay type
Cell-based (qualitative)
Reactive species
Mammals

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Target data

Function

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).

Alternative names

What's included?

500 Test
Components
Blue Fluorescent Phalloidin Conjugate
1 x 1 Vial
Labeling Buffer
1 x 50 mL

Recommended products

F-actin Staining Kit - Blue Fluorescence | Cytopainter (ab112124) fluorescence imaging kits are a set of fluorescence imaging tools for labeling sub-cellular organelles such as lysosomes, mitochondria, and actin filaments.

Key facts

Sample types
Tissue, Suspension cells, Adherent cells
Assay type
Cell-based (qualitative)
Reactive species
Mammals
Assay Platform
Fluorescence microscope

Storage

Shipped at conditions
Blue Ice
Appropriate short-term storage conditions
-20°C
Appropriate long-term storage conditions
-20°C
Storage information
-20°C

Notes

F-actin Staining Kit - Blue Fluorescence | Cytopainter (ab112124) fluorescence imaging kits are a set of fluorescence imaging tools for labeling sub-cellular organelles such as lysosomes, mitochondria, and actin filaments. The selective staining of cell compartments provides a powerful method for studying cellular events in a spatial and temporal context.

ab112124 is designed to stain F-actins in fixed cells with blue fluorescence. The kit uses a blue fluorescent phalloidin conjugate that is selectively bound to F-actins. The phalloidin conjugate has Ex/Em = 350/450 nm, compatible with DAPI filter set that comes with most of fluorescence microscopes. It is a high-affinity probe for labeling, identifying and quantitating F-actins in formaldehyde-fixed and permeabilized tissue sections, cell cultures or cell-free experiments. ab112124 provides all the essential components with an optimized labeling protocol.

Store ab112124 desiccated.

Supplementary info

This supplementary information is collated from multiple sources and compiled automatically.
Activity summary

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.

Biological function summary

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.

Pathways

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.

Associated diseases and disorders

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.

Product promise

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In the unlikely event of one of our products not working as expected, you are covered by our product promise.

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2 product images

  • Fluorescence Microscopy - F-actin Staining Kit - Blue Fluorescence - Cytopainter (ab112124), expandable thumbnail

    Fluorescence Microscopy - F-actin Staining Kit - Blue Fluorescence - Cytopainter (ab112124)

    Image: Courtesy of Dr. Shaohua Li, UMDNJ-Robert Wood Johnson Medical School

    Sample: mouse embryonic stem cell-differentiated embryoid bodies (EBs)


    Preparation:

    • Fix in 3%PFA in PBS for 30 min at RT
    • Incubate in 7.5% sucrose-PBS for 3h at RT
    • Incubate in 15% sucrose-PBS at 4 degree Celsius overnight
    • Embed the EBs in tissue-Tek OCT compound
    • Cut frozen sections to 4-20 µm thickness

    Primary antibody: Rabbit anti laminin alpha 1, 1:400

    Secondary antibody: Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Cy5®) (Goat Anti-Rabbit IgG H&L (Cy5 ®) preadsorbed ab97077), 1:100

    F-actin was stained with CytoPainter blue (ab112124), 1:100

    Nuclei were counterstained with SYTO 16

  • Fluorescence Microscopy - F-actin Staining Kit - Blue Fluorescence - Cytopainter (ab112124), expandable thumbnail

    Fluorescence Microscopy - F-actin Staining Kit - Blue Fluorescence - Cytopainter (ab112124)

    CytoPainter F-actin Labelling Kit - Blue Fluorescence (ab112124) - CPA cells were fixed with 4.0% formaldehyde in a Costar black 96-well plate.
    Left panel: Cells were labelled with CytoPainter F-actin Labelling Kit - Blue for 30 minutes. Right panel: Cells were pre-treated for 10 minutes with phalloidin, prior staining with CytoPainter F-actin Labelling Kit - Blue for 30 minutes.

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Product protocols

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