Overview

  • Product name
    F-actin Staining Kit - Red Fluorescence - Cytopainter
    See all F-actin kits
  • Sample type
    Tissue, Adherent cells, Suspension cells
  • Assay type
    Cell-based
  • Species reactivity
    Reacts with: Mammals, Other species
  • Product overview

    F-actin Staining Kit - Red Fluorescence | Cytopainter (ab112127) 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.


    ab112127 is designed to stain F-actins in fixed cells with red fluorescence. The red fluorescent phalloidin conjugate, which is selectively bound to F-actins, is a high-affinity probe for F-actins. The red fluorescent phalloidin conjugate has Ex/Em = 594/610 nm. Used at nanomolar concentrations, phallotoxins can be conveniently used to label, identify and quantitate F-actins in formaldehyde-fixed and permeabilized tissue sections, cell cultures or cell-free experiments. The red fluorescent phalloidin conjugate has good thermal and photo stability.


    ab112127 provides all the essential components with an optimized labeling protocol. It is an excellent tool for preserving fluorescent images of particular cells, and can also be used for fluorescence microscope demonstrations.

  • Notes

    ab112127 should be stored dessicated.

  • Platform
    Fluorescence microscope

Properties

Images

  •  F-actin staining (red) in chondrocytes in growth plate cartilage. Mouse femur bone was fixed with 4% PFA for 72 hours, decalcified and cut into 8-10 µm sections in frozen. Tissue was then stained for 60 minutes with ab112127 before a final wash in PBS.

     

    This image is courtesy of an anonymous Abreview

  • HeLa cells were fixed with 4% PFA for 10 minutes, rinsed with PBS and stained for 60 minutes with ab112127 before a final wash in PBS prior to mounting slides.

    Images obtained with an Olympus BX61 microscope using a Texas Red filter (595/613nm) - 50ms exposure.

  • Images of CPA cells fixed with formaldehyde and stained with ab112127 in a black 96-well plate Left image: Cells labeled with 1X Red Fluorescent Phalloidin Conjugate for 30 min only. Right image: Cells treated with phalloidin for 10 min, then stained with Red Fluorescent Phalloidin Conjugate for 30 min.

Protocols

References

This product has been referenced in:
  • Toprakhisar B  et al. Development of Bioink from Decellularized Tendon Extracellular Matrix for 3D Bioprinting. Macromol Biosci N/A:e1800024 (2018). Read more (PubMed: 30019414) »
  • Topuz F  et al. Nanosilicate embedded agarose hydrogels with improved bioactivity. Carbohydr Polym 201:105-112 (2018). Read more (PubMed: 30241801) »
See all 4 Publications for this product

Customer reviews and Q&As

1-10 of 12 Abreviews or Q&A

CytoPainter F-actin Staining Kit for IHC-P

Poor Excellent 5/5 (Ease of Use)
Abreviews
I have tried this Kit on paraffin embedded sections of adult human kidney. Because the protocol is for ICC I adjusted it to IHC. In addition, I combined it with HLA staining (ab52922, routinely used at our lab) to visualize the tissue.

Deparaffinization and antigen retrieval was done with commercial - ZYTOMED's OmniPrep (pH 9). Slides were washed with PBS-Tween (0.05%) and blocked (commercial - Invitrogen's CAS-Block) – 1h at RT. Anti-HLA was incubated for 1h at RT and washed with PBST three times.

Phalloidin reagent was prepared according to protocol in three dilutions: 1/250, 1/500 and 1/1000. Secondary Alexa flour 488 donkey anti rabbit was added to each reagent tube at a 1/1200 dilution. Slides were incubated for 1h at RT and then washed three times with PBST. Secondary only slices (only Secondary Alexa flour 488) were used to determine exposure time threshold.
There was no signal above background in all three Phalloidin dilutions. The picture (at the Texas Red Tfilter) was taken in slight overexposure to show the absent of staining.

Abcam user community

Verified customer

Submitted Jul 10 2017

Cytopainter F-actin in Hydractinia

Excellent Excellent 5/5 (Ease of Use)
Abreviews
We used cytopainter F-actin staining kit to study morphology in the model organism Hydractinia during head regeneration. We fixed our whole animals in 4% pararfomaldehde in PBS followed by three ten minute washes in 0.1%triton in PBS. The animals were incubated in component in at a 1:500 dilution in component B for 45 minutes. They were then washed in PBS three times and mounted in fluorescent mounting medium.
The picture shows F-actin staining(Red) and Hoescht nuclear staining(blue). The image was taken on a spinning disc confocal microscope.

Mr. James Gahan

Verified customer

Submitted Jan 23 2014

Answer

Yes, I believe they are also in the bone matrix.

http://www.ncbi.nlm.nih.gov/pubmed/9610818.

Read More

Answer

Yes, it does. Please see attached reference.





http://www.ncbi.nlm.nih.gov/pubmed/9610818



http://cjasn.asnjournals.org/content/3/Supplement_3/S131.full.pdf+html

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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.

Read More

Answer

Thank you for contacting us.

This kit has an excitation of 583 nm and an emission at 603 nm. I have contacted the lab who confirm that this kit can be used with other antibodies providing that its emission does not overlap with the kits emission.

I recommend using an Alexa488 conjugated secondary antibody against the primary antibody that you wish to use in conjugation with this kit.

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 confirming these details and for your cooperation. The details provided enable us to closely monitor the quality of our products.

I am sorry this product did not perform as stated on the datasheet and for the inconvenience this has caused. As requested, I have issued a free of charge replacement with the order number ***.

To check the status of the order please contact our Customer Service team and reference this number.

Please note that this free of charge replacement vial is also covered by our Abpromise guarantee. Should you still be experiencing difficulties, or if you have any further questions, please do not hesitate to let us know.

I wish you the best of luck with your research.

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Answer

Thank you for your reply and I am sorry about the confusion over the component list and the protocol.

As you have added the Methanol to the already reconstituted Component A, I would be happy to send you a new kit, as I am sure if the Component A of the kit you already have is viable. To be able to send you the new kit, would you please be able to provide me with either the Abcam order number, or the PO number that was used to purchase the antibody? Once I receive that information, I will be happy to get you a replacement kit as soon as I can.

I look forward to your reply.

Read More

Answer

Thank you for contacting Abcam.

I am sorry about the mix up, but it seems as though you received the older version of the protocol. Also, in the newer version of the kit, there is only Component A & B, and component A is already in a liquid form. I have also attached the up-to-date protocol.

If there is anything else I can help you with, please let me know.

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Answer

Vielen Dank für Ihren Anruf.

Ich freue mich Ihnen bestätigen zu können, dass die 50x Stock Solution von Red Fluorescent Phalloidin Conjugate für sechs Monate bei -20C aufgehoben werden kann.

Ich hoffe diese Information ist hilfreich und wünsche viel Erfolg mit Ihren Experimenten.

Es war schön Sie bei unserer Roadshow kennenzulernen!

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1-10 of 12 Abreviews or Q&A

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