All plasmids used in this study are available from Addgene (Cambridge, MA). mCherry-Alpha-Actinin2-N-18 (Addgene plasmid #54974), mApple-Lifeact-7 (Addgene plasmid #54747), mEmerald-Lifeact-7 (Addgene plasmid #54148), mCherry-Paxillin-22 (Addgene plasmid #55114) (Paszek et al., 2012 (link)), EGFP-Talin-H-18 (Addgene plasmid #56448), and EGFP-Talin-CW-18 (Addgene plasmid # 56446) were gifts from Michael Davidson.
Memerald lifeact 7
Manufactured by Addgene
MEmerald-Lifeact-7 is a fluorescent protein marker that specifically labels actin filaments in live cells. It is a fusion of the green fluorescent protein mEmerald and the actin-binding peptide Lifeact-7, which allows for real-time visualization of the actin cytoskeleton.
Automatically generated - may contain errors
Lab products found in correlation
Mapple lifeact 7, by Addgene (2 mentions)
Lipofectamine 3000 transfection reagent, by Thermo Fisher Scientific (1 mentions)
Alexa fluor 568 protein labeling kit, by Thermo Fisher Scientific (1 mentions)
Gfr matrigel, by BD (1 mentions)
Latrunculin b, by Merck Group (1 mentions)
Dq collagen, by Thermo Fisher Scientific (1 mentions)
Purecol ez gel, by Advanced BioMatrix (1 mentions)
Alexa fluor 488 phalloidin, by Thermo Fisher Scientific (1 mentions)
4 6 diamidino 2 phenylindole (dapi), by Thermo Fisher Scientific (1 mentions)
Dimethyl sulfoxide (dmso), by Merck Group (1 mentions)
Pspax2, by Addgene (1 mentions)
Pmd2 g, by Addgene (1 mentions)
4 protocols using memerald lifeact 7
1
Plasmid Library for Cell Adhesion Studies
2
Fluorescent Protein Transfection
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Plasmids were purchased from Addgene: mEmerald-Lifeact-7 (54148) and pmEGFP-α-tubulin-IRES-puro2b (21042). pSems-ClathrinLC-Halotag plasmid was a gift from J. Piehler (University of Osnabrück). Transfection was performed with Lipofectamine 3000 Transfection Reagent (Invitrogen) according to the manufacturer’s protocol.
3
Immunofluorescence Microscopy with Cytoskeletal Markers
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The anti-MAP2 antibody was acquired from Antibodies Online (ABIN372661; used at 1:1,000 for immunofluorescence). The Alexa Fluor 488 Phalloidin (A12379), used to stain filamentous actin, and DAPI (D1306) were purchased from Life Technologies. Bovine plasma FN was purchased from Merck (341631). DMSO and latrunculin B (L5288-1MG) were obtained from Sigma-Aldrich. GFR Matrigel was bought from BD Biosciences (354230). PureCol EZ Gel (fibrillar collagen I, concentration 5 mg/ml) was provided by Advanced BioMatrix. DQ collagen (type I collagen from bovine skin, fluorescein conjugate; D12060 ) was provided by Thermo Fisher Scientific. mEmerald-Lifeact-7 was a gift from M. Davidson (plasmid 54148; Addgene). psPAX2 and pMD2.G were gifts from D. Trono (École polytechnique fédérale de Lausanne, Lausanne, Switzerland; plasmids 12260 and 12259; Addgene). pCDH-LifeAct-mRFP was a gift from P. Caswell. Full-length bovine FN was labeled with Alexa Fluor 568 using an Alexa Fluor 568 Protein Labeling kit (A10238 ; Thermo Fisher Scientific) according to the manufacturer’s instructions.
4
Lifeact-Labeled Actin Dynamics
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