Icahn School of Medicine at Mount Sinai LINCS Center for Drug Toxicity Signatures

Immunofluorescence in undifferentiated PromoCell cardiomyocytes (Line B) shows only diffuse cytoplasmic expression of sarcomeric actinin (green) with little or no co-localization with the actin cytoskeleton (red).

Immunofluorescence in PromoCell cardiomycytes (Line B) differentiated for 4 weeks shows increased focal expression of sarcomeric actinin (green) that strongly co-localizes with adhesions sites and the actin cytoskeleton (red).

Assay Date(s): 08/11/2015
Primary Antibody: Mouse monoclonal anti-alpha-actinin (sarcomeric)
(Sigma, Cat: A7811-100UL, Lot: 103M4770)
Secondary Antibody: Alexa Fluor 488 goat anti-mouse
(Invitrogen, Cat: A11001, Lot: 1397999)
Temperature: 37ºC
Dilution 1:100
Diluent: 10% goat serum, 2.5% bovine serum albumin in PBS
Actin Staining: Alexa Fluor 647 phalloidin
(Invitrogen, Cat: A22287, Lot: 648322) )
Dilution: 1:40
Diluent: PBS
Nuclear Staining: Hoechst 33342, trihydrochloride, trihydtrate
(Invitrogen, Cat: H3570, Lot: 46C3-1)
Dilution: 1:10,000
Diluent: PBS
Imaging Date: 08/11/2015
Imaging Platform: Zeiss LSM 780, Axio Imager 2
Objective: Plan-Apo 40x/1.4 Oil DIC M27
Excitation: 458 (Argon), 633 (HeNe 633), 405 (Diode 405-30)
Emission: 410-508, 490-632, 638-755
Related Standard Operating Procedures (SOPs)
Cell SOPs: DToxS SOP CE – 1.0: PromoCell Cardiomyocyte Subculture
DToxS SOP CE – 2.0: PromoCell Cardiomyocyte Plating for Drug Test
DToxS SOP CE – 4.0: Drug Treatment and Cell Lysis
Assay SOPs: DToxS SOP A – 4.0 Immunofluorescence PromoCell Cardiomyocytes
Performed by: B. Hu, G. Jayaraman, R. C. Calizo

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