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: Other species, Mammals
  • 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.

  • Platform

    Fluorescence microscope

Properties

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

This product has been referenced in:

  • Song Y  et al. Epithelial C5aR1 Signaling Enhances Uropathogenic Escherichia coli Adhesion to Human Renal Tubular Epithelial Cells. Front Immunol 9:949 (2018). Read more (PubMed: 29765378) »
See 1 Publication for this product

Customer reviews and Q&As

1-8 of 8 Abreviews or Q&A

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.

Abcam user community

Verified customer

Submitted Aug 09 2017

F-actin staining in rat hepatocyte - blue - F-actin green - trans-Golgi staining by TGN46

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.

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.

Prof. Martin Mendelson

Verified customer

Submitted Mar 17 2014

Question
Answer

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 providing that their emissions do not overlap with the kit's emission.

https://www.abcam.com/index.html?pageconfig=resource&rid=14270

I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

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Question
Answer

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 twice in PBS.
To help expose the insoluble beta Amyloid, we recommend performing antigen retrieval with 10% formic acid in distilled water, pH 1.6-2.0 for 10-30 minutes at room temperature.
Incubate sections for1 hourin 10% normal serum from species in which secondary antibody was raised. Tap excess serum off the slides before staining.
Incubate sections in primary antibody for at least 1 hour at room temperature in a humid chamber or overnight at 4°C. For ab10148, we recommend a starting dilution between 1:100-1:500, and a range of 1:300-1:2000 for ab110888.Wash three times in PBS.
Addfluorescentsecondary antibody at recommended dilution (see specific datasheet for details). Incubate for at least 30 minutes at room temperature in the dark. Wash three times in PBS.
Counterstain with DAPI or our CytoPainter dyes: https://www.abcam.com/CytoPainter
Mount in aqueous mounting medium.



I hope this helps, please let me know if you need any other information or assistance.

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Question
Answer

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

https://www.abcam.com/CytoPainter


I hope this helps, please let me know if you have any other questions and I will be happy to help you further.

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Question
Answer

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.

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