F-actin Staining Kit - Green Fluorescence - Cytopainter (ab112125)

Abreviews (1)Q&A (4)Specific References (1)

Overview

  • Product name
    F-actin Staining Kit - Green Fluorescence - Cytopainter
    See all F-actin kits
  • Sample type
    Tissue, Adherent cells, Suspension cells
  • Assay type
    Cell-based (qualitative)
  • Product overview

    F-actin Staining Kit - Green Fluorescence | Cytopainter (ab112125) 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.


    ab112125 is designed to stain F-actins in fixed cells with green fluorescence. The kit uses a green fluorescent phalloidin conjugate that is selectively bound to F-actins. This green fluorescent phalloidin conjugate is a high-affinity probe for F-actins. When used at nanomolar concentrations, phallotoxins are convenient probes for labeling, identifying and quantitating F-actins in formaldehyde-fixed and permeabilized tissue sections, cell cultures or cell-free experiments.


    ab112125 provides all the essential components with an optimized staining protocol, which is robust requiring minimal hands-on time. It is an excellent tool for preserving the fluorescent images of particular cells. The phalloidin conjugate has spectral properties similar to those of FITC (Ex/Em = 500/520 nm). It can also be used for fluorescence microscope demonstrations using FITC filter (Ex/Em = 490/525 nm).


     

  • Notes

    ab112125 should be stored dessicated.

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

Properties

Applications

Our Abpromise guarantee covers the use of ab112125 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.
ICC/IF Use at an assay dependent concentration.

Images

  • Functional Studies - CytoPainter F-actin Labeling Kit - Green Fluorescence (ab112125)
    Functional Studies - CytoPainter F-actin Labeling Kit - Green Fluorescence (ab112125)
    Left Panel: Cells labeled with 1X Green Fluorescent Phalloidin Conjugate for 30 minutes only.
    Right Panel: Cells treated with phalloidin for 10 minutes, then stained with Green Fluorescent Phalloidin Conjugate for 30 minutes.

Protocols

References

This product has been referenced in:
  • Banni M  et al. Assessing the impact of Benzo[a]pyrene on Marine Mussels: Application of a novel targeted low density microarray complementing classical biomarker responses. PLoS One 12:e0178460 (2017). Read more (PubMed: 28651000) »

See 1 Publication for this product

Publishing research using ab112125? Please let us know so that we can cite the reference in this datasheet.

Customer reviews and Q&As

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5 reviews or Q&A

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Votes: Highest First

Cytopainter F-actin Staining Kit - Green Fluorescence for STED microscopy

 Average Good 4/5 (Ease of Use)
Abreviews
The kit is easy to use. This is an advantage, though the fact that it comes with a buffer to use for the fixation protocol does not allow for easy versatility. Is a bit dis courageous to try using it adding other fluoroscopes and knowing which buffer suits the other fluorophores best.
For STED purposes, it could be used, but we found a bit higher the background noise. This might be as well the buffer, we did not test it further.
We used not very high STED power and work, but not being better than other dyes we are used to employ for actin staining.
abreview image
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Verified customer

Submitted Mar 03 2015

Tissues are dissected in 1x PBS.

Samples are then immersed in 4% PFA in PBS overnight at 4°C

This is followed by dehydration via methanol replacement (25%, 50%, 75%, 100%x3). 1 hour per increase in % methanol. Samples are kept...

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Thank you for your protocol information.



Using methanol to dehydrate may be causing damage to the actin stucture. Please consider using the following ethanol dehydration protocol.



Fixation: 4% PFA in PBS overni...

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I've tried CytoPainter on my paraffin embedded samples, but it didn't work probably owing to the methanol dehydration step after PFA fixation during the preparation for embedding.

To my knowledge, dehydration would be required since paraffin...

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Could you please provide your fixation and embedding protocols? As fas as I know, I have not come across any protocols that required methanol dehydration steps. Hopefully with this information, I will be able to help troubleshoot your experiment.

One more thing, should the 1x green fluorescent phallodin working solution always be prepared fresh? Would it be possible to store the excess 1x working solution at -20C for later use?

Thank you for your enquiry.



It is best to prepare fresh solution, so if you do not need to use 10ml in one experiment, then you could start with making less working solution (eg 5ul conjugate in 5mL buffer). Please avoid aliquating...

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Inquiry: Cytopainter F-actin Staining Kit - Green Fluorescence (ab112125) Will the antigen retrieval step be required for this product to work on paraffin embedded murine samples? Also, can this be used in conjunction with other antibodies to produce d...

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Thank you for your enquiry.



Here are the answers to your questions.



1) Antigen retrieval will be required if you have used PFA or formalin to fix your tissue. I would suggest trying a section that has been retr...

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

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