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

Overview

  • Product name
    F-actin Staining Kit - Blue Fluorescence - Cytopainter
    See all F-actin kits
  • Sample type
    Tissue, Adherent cells, Suspension cells
  • Assay type
    Cell-based (qualitative)
  • Species reactivity
    Reacts with: Human
  • Product overview

    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.


     

  • Notes

    Store ab112124 desiccated.

  • Tested applications
    Suitable for: FM, ICC/IFmore details

Properties

Applications

Our Abpromise guarantee covers the use of ab112124 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
FM Use at an assay dependent concentration.

Use formaldehyde-fixed and permeabilized tissue sections or cell culture.

ICC/IF Use at an assay dependent concentration.

Use formaldehyde-fixed and permeabilized tissue sections or cell culture.

Images

  • 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®) (ab97077), 1:100
    F-actin was stained with CytoPainter blue (ab112124), 1:100
    Nuclei were counterstained with SYTO 16

  • 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.

Protocols

References

ab112124 has not yet been referenced specifically in any publications.

Customer reviews and Q&As

Abreviews
The kit is straight forward and works well. However, you have to consider the fixation method. I used 4% PFA and perhaps, is not the best method for visualizing component of the cytoskeleton.
Additionally, I strongly suggest adjusting the dilution of the Blue Phalloidin in the provided buffer because highly dependent on the experimental conditions and therefore, not universally valid.

The picture shows murine derived breast cancer cells (untreated) stained for F-actin.
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Abcam user community

Verified customer

Submitted Aug 09 2017

F-actin staining in rat hepatocyte - blue - F-actin green - trans-Golgi staining by TGN46
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Dr. Armen Petrosyan

Verified customer

Submitted Oct 22 2015

F-actin of human HEPG-2 cells have been stained using CytoPainter F-actin Staining Kit-Blue Fluorescence according to the manufacture’s (Abcam) protocol.
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Abcam user community

Verified customer

Submitted Aug 19 2015

Actin in crustacean mechanoreceptors

Good Excellent 5/5 (Ease of Use)
Abreviews
Good bright blue signal with easy procedure. Interestingly, there seems to be some differentiation as to which structures are visualized depending on which detergent is used for permeabilization. This needs to be pursued.
My only reservation is that fading over time at 4ºC has been more notable than with the Alexafluors.
Username

Dr. Martin Mendelson

Verified customer

Submitted Mar 17 2014

Thank you for contacting us.

The CytoPainter kits are compatible with secondary antibodies and nuclear dyes for multicolor staining or GFP-transfected cells, for example. The lab has confirmed that these kits can be used with other antibodies...

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Thank you for contacting us. To perform IHC-P with fluorescent secondary antibodies, I would recommend the following protocol:

De-paraffinise sections thoroughly in Xylene/synthetic solvent and hydrate through graded series of alcohols. Wash ...

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Thank you for your phone call. The dyes we were discussing are available in several colors, and the full list can be found here:

http://www.abcam.com/CytoPainter


I hope this helps, please let me know if you have any other quest...

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Thank you for your enquiry. I have looked into this issue and ab112124 does only stain dead (fixed) cells. Sorry for the confusion. Please feel free to contact me with any further questions.

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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