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Acti stain 670

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Acti-stain 670 is a fluorescent dye that specifically labels actin filaments in cells. It can be used to visualize the actin cytoskeleton in a variety of cell types.

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Lab products found in correlation

Rhodamine phalloidin, by Thermo Fisher Scientific (6 mentions) Lamp1 bs 1970r, by Bioss Antibodies (2 mentions) Hic 5 611164, by BD (2 mentions) β1 integrin clone 9eg7 553715, by BD (2 mentions) Cd29 610467, by BD (2 mentions) Ilk 611803, by BD (2 mentions) Tensin1 nbp1 84129, by Novus Biologicals (2 mentions) Phosphotyrosine clone 4g10 05 321, by Merck Group (2 mentions) Talin t3287, by Merck Group (2 mentions) α actinin a5044, by Merck Group (2 mentions) Tensin1 sab200283, by Merck Group (2 mentions) Phosphotyrosine clone p tyr 100 9411, by Cell Signaling Technology (2 mentions) β1 integrin clone 12g10 ab30394, by Abcam (2 mentions) α tubulin clone dm1a t9026, by Merck Group (2 mentions) Fibronectin f3648, by Merck Group (2 mentions) Anti rabbit secondary antibody, by Jackson ImmunoResearch (2 mentions) Pcsdest vector, by Addgene (1 mentions) Mmessage machine sp6 kit, by Thermo Fisher Scientific (1 mentions) Evos xl core imaging system, by Thermo Fisher Scientific (1 mentions) Sp8 inverted confocal microscope, by Leica (1 mentions)

Spelling variants of this product

Acti-stain 670 phalloidin (13 citations) Acti-stain (7 citations)

6 protocols using acti stain 670

1

Fluorescent Rac1 Visualization in Cells

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HeLa or COS7 cells were transiently transfected with plasmid encoding CFP-Rac1(CA). Twelve hours later, cells were fixed with 4% P FA. After permeabilization with PBS containing 0.5% triton X-100 for 5 min, cellular actin was detected with Acti-stain 670 phalloidin (Cytoskeleton).
Onuma H., Komatsu T., Arita M., Hanaoka K., Ueno T., Terai T., Nagano T, & Inoue T. (2014). Rapidly rendering cells phagocytic through a cell-surface display technique and concurrent Rac activation. Science signaling, 7(334), rs4.
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2

Integrin and Cytoskeletal Protein Analysis

Cited 1 time
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Antibodies used in this analysis were: Hic-5 611164, β1 integrin clone 9EG7 553715, CD29 610467, ILK 611803 (BD Biosciences, Franklin Lakes, NJ USA); tensin1 NBP1-84129 (Novus Biologicals, Littleton, CO, USA); phosphotyrosine clone 4G10 05-321 (Millipore); talin T3287, fibronectin F3648, α-tubulin clone DM1A T9026, α-actinin A5044, tensin1 SAB200283 (Sigma-Aldrich, St. Louis, MO, USA); phosphotyrosine clone P-Tyr-100 9411 (Cell Signaling, Danvers, MA, USA); β1 integrin clone 12G10 ab30394 (Abcam, Cambridge, MA, USA); LAMP1 bs-1970R (Bioss, Woburn, MA, USA); Epcam G8.8 was deposited to the DSHB by Farr, A.G.
F-actin was visualized using Rhodamine phalloidin (Thermo-Fisher, Carlsbad, CA, USA) or Actistain 670 (Cytoskeleton, Denver, CO, USA). Fluorescently conjugated secondary antibodies used were: Dylight 488, 550 and 633 conjugated anti-mouse and anti-rabbit were purchased from Thermo-Fisher. FITC anti-rat, Alexa-fluor 488 anti-mouse, HRP-conjugated anti-mouse and anti-rabbit secondary antibodies were purchased from Jackson ImmunoResearch. The polydimethylsiloxane (PDMS) substrates were prepared as previously described(52 (link)).
Goreczny G.J., Forsythe I.J, & Turner C.E. (2018). Hic-5 Regulates Fibrillar Adhesion Formation to Control Tumor Extracellular Matrix Remodeling through Interaction with Tensin1. Oncogene, 37(13), 1699-1713.
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3

Integrin and Cytoskeletal Protein Analysis

Cited 1 time
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Antibodies used in this analysis were: Hic-5 611164, β1 integrin clone 9EG7 553715, CD29 610467, ILK 611803 (BD Biosciences, Franklin Lakes, NJ USA); tensin1 NBP1-84129 (Novus Biologicals, Littleton, CO, USA); phosphotyrosine clone 4G10 05-321 (Millipore); talin T3287, fibronectin F3648, α-tubulin clone DM1A T9026, α-actinin A5044, tensin1 SAB200283 (Sigma-Aldrich, St. Louis, MO, USA); phosphotyrosine clone P-Tyr-100 9411 (Cell Signaling, Danvers, MA, USA); β1 integrin clone 12G10 ab30394 (Abcam, Cambridge, MA, USA); LAMP1 bs-1970R (Bioss, Woburn, MA, USA); Epcam G8.8 was deposited to the DSHB by Farr, A.G.
F-actin was visualized using Rhodamine phalloidin (Thermo-Fisher, Carlsbad, CA, USA) or Actistain 670 (Cytoskeleton, Denver, CO, USA). Fluorescently conjugated secondary antibodies used were: Dylight 488, 550 and 633 conjugated anti-mouse and anti-rabbit were purchased from Thermo-Fisher. FITC anti-rat, Alexa-fluor 488 anti-mouse, HRP-conjugated anti-mouse and anti-rabbit secondary antibodies were purchased from Jackson ImmunoResearch. The polydimethylsiloxane (PDMS) substrates were prepared as previously described(52 (link)).
Goreczny G.J., Forsythe I.J, & Turner C.E. (2018). Hic-5 Regulates Fibrillar Adhesion Formation to Control Tumor Extracellular Matrix Remodeling through Interaction with Tensin1. Oncogene, 37(13), 1699-1713.
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4

Overexpression of ACTA1 Mutants

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Wild-type and Glu197Asp human ACTA1 genes were cloned into pCSDest vector (Addgene) as described.15 (link) Plasmids were linearized with restriction enzyme NotI, and RNA was transcribed using a kit (SP6mMessage Machine kit; Ambion). One-cell stage embryos were injected with 200 pg and 400 pg of RNA, and muscle was analyzedas described previously16 (link) using Acti-stain 670 (Cytoskeleton Inc) and antimyosin heavy chain antibody (F59; Santa Cruz Biotech).
Zukosky K., Meilleur K., Traynor B.J., Dastgir J., Medne L., Devoto M., Collins J., Rooney J., Zou Y., Yang M.L., Gibbs J.R., Meier M., Stetefeld J., Finkel R.S., Schessl J., Elman L., Felice K., Ferguson T.A., Ceyhan-Birsoy O., Beggs A.H., Tennekoon G., Johnson J.O, & Bönnemann C.G. (2015). Association of a Novel ACTA1 Mutation With a Dominant Progressive Scapuloperoneal Myopathy in an Extended Family. JAMA neurology, 72(6), 689-698.
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5

Multicolor Immunofluorescence Imaging

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Alexa Fluor 488 Donkey anti-mouse (1:500; ThermoFisher, A21202), Alexa Fluor 555 Donkey anti-rabbit (1:500; ThermoFisher, A32727), Alexa Fluor 647 Donkey anti-goat (1:500, ThermoFisher, A21447)
F-actin staining: Acti-Stain 670 (1:100; Cytoskeleton, Inc., PHDN1-A)
Immunofluorescence images were acquired on an inverted SP8 confocal microscope (Leica Microsystems) with the 20x/0.75 IMM CS2 (HC PL APO) objective with a z-stack of 2 μm. Image preparation was conducted in Fiji software69 (link). Laser power and detector gain were maintained constant within a single experiment.
Brightfield images were acquired on an EVOS XL Core Imaging System (AMEX1100, ThermoFisher).
Bergmann S., Penfold C.A., Slatery E., Siriwardena D., Drummer C., Clark S., Strawbridge S.E., Kishimoto K., Vickers A., Tewary M., Kohler T.N., Hollfelder F., Reik W., Sasaki E., Behr R, & Boroviak T.E. (2022). Spatial profiling of early primate gastrulation in utero. Nature, 609(7925), 136-143.
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6

Visualizing Acetylcholine Receptors and Cytoskeleton

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The cells were fixed in 4% paraformaldehyde (PFA) and stained using AlexaFluor-555-bugarotoxin (BTX) (catalog no. B35451, Life Technologies) and Alexa-488-taged BTX (catalog no. B13422, Life Technologies). For labelling of AChRs in live cells BTX conjugates were added to the culture media for 5–7 min; internal AChR staining was performed after cell membrane permabilization with 0.5% Triton X-100 for 1 h. For masking of BTX binding sites on surface AChRs unlabelled BTX (catalog no. B1601, Life Technologies) was added to the culture media for 20 minutes. For microtubule disruption 10 µg/ml nocodazole (catalog no. M1404, Sigma) or DMSO (control) was added to the media for 6 hours. Microtubules were visualised with anti-tubulin (catalog no. ab18251, Abcam) and EB1 with anti-EB1 (catalog no. 610534, BD Biosciences). F-actin was stained with Acti-stain-670 (catalog no. PHDN1, Cytoskeleton).
Bernadzki K.M., Gawor M., Pęziński M., Mazurek P., Niewiadomski P., Rędowicz M.J, & Prószyński T.J. (2017). Liprin-α-1 is a novel component of the murine neuromuscular junction and is involved in the organization of the postsynaptic machinery. Scientific Reports, 7, 9116.
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