Acti-stain™ Fluorescent Phalloidins

The Acti-stain™ line of fluorescent phalloidins has been developed with an emphasis on creating exceptionally bright and stable probes at an economical price. Side-by-side comparisons with leading competing products demonstrate that you will enjoy savings while not sacrificing the quality of the stain when switching to an Acti-stain™ probe. Additionally, these probes are compatible with popular filter sets such as FITC, TRITC and Cy 5. The combination of in-house manufacturing, stringent quality control and convenient packaging provides a great value. Give them a try and see for yourself.

Product Uses Include

  • Stain F-actin in fixed cells
  • Stabilize actin filaments in vitro
  • Visualize actin filaments in vitro 

Actin staining is very useful in determining the structure and function of the cytoskeleton in living and fixed cells. The actin cytoskeleton is a very dynamic and labile structure in the living cell, but it can be fixed with paraformaldehyde prior to probing or staining for actin structures.

Click here for Acti-stain specifications

Click here for General F-actin staining information and protocols

PHDG1_img3

Cytoskeleton's Acti-stain products have been cited many times since their introduction, here are the most recent examples:

Product Citations for Acti-stain fluorescent probes

AuthorTitleJournalYearArticle Link
Takaya, Kento et al.Salicylate induces epithelial actin reorganization via activation of the AMP-activated protein kinase and promotes wound healing and contraction in miceScientific Reports2024
Wang, Chunqing et al.PKCζ phosphorylates VASP to mediate chemotaxis in breast cancer cellsExperimental Cell Research2023
Phillips, Andrew T. et al.The formin DAAM1 regulates the deubiquitinase activity of USP10 and integrin homeostasisEuropean Journal of Cell Biology2023
Boonrod, Kajohn et al.Identification of the Actin-Binding Region and Binding to Host Plant Apple Actin of Immunodominant Transmembrane Protein of ‘Candidatus Phytoplasma mali’International Journal of Molecular Sciences2023
Chang, Yongyun et al.Aspirin prevents estrogen deficiency-induced bone loss by inhibiting osteoclastogenesis and promoting osteogenesisJournal of Orthopaedic Surgery and Research2023
Bhosle, Vikrant K. et al.The chemorepellent, SLIT2, bolsters innate immunity against Staphylococcus aureuseLife2023
Karvas, Rowan M. et al.3D-cultured blastoids model human embryogenesis from pre-implantation to early gastrulation stagesCell stem cell2023
Park, Jinho et al.Graphene Enhances Actin Filament Assembly Kinetics and Modulates NIH-3T3 Fibroblast Cell SpreadingInternational Journal of Molecular Sciences2022

Question 1:  Which is the most stable/brightest Acti-stain conjugate? 

Answer 1: The brightest and most stable of the Acti-stains is Acti-stain 488 (green fluorescence; Cat. # PHDG1).  Please see the table below for additional information on all of our Acti-stains. 

Conjugate

Cat. #

Wavelengths (Ex/EM)

Brightness (AFU)

Stability to photobleaching* (1/2 life in sec)

Background (% of total AFU at 100 nM)

Acti-stain™ 488

PHDG1

485/535

832

57

5

Acti-stain™ 535

PHDR1

535/585

430

27

12

Acti-stain™ 555

PHDH1

535/585

551

46

16

Acti-stain™ 670

PHDN1

640/680

837

38

12

* = measured in the absence of antifade.

Question 2:  What fixation conditions are recommended when staining with phalloidin conjugates? 

Answer 2: Fluorescent phalloidins only bind to the native quaternary structure of F-actin which provides a low background.  The correct fixation condition for phalloidin binding is 3.7% (v/v) paraformaldehyde in PBS for 10 min because it retains the quaternary protein structure which is necessary for high affinity binding of phalloidin.  Methanol fixation destroys the native conformation and hence is not suitable for F-actin staining with phalloidin.

For more information about staining procedures and specifications of the Acti-stain series, please click here.