3×105 cells were adhered onto polylysine D-coated coverslips (Sigma Aldrich) during a 45-minute incubation at 37°C, fixed in 10% formalin (Fisher Scientific, Pittsburgh, PA) and permebealized in 1% triton-X 100 in PBS. Coverslips were blocked for 30 minutes in 5% bovine serum albumin (Sigma-Aldrich) in PBS with 0.1% Tween-20, probed with p65/RelA, p52/p100 (Cell Signaling) or cleaved PARP (Thermo Scientific, Lafayette, CO) antibodies and then with Alexa Fluor 594 goat anti-mouse antibodies (Life Technologies). Coverslips were mounted with anti-fading ProLong Gold Solution (Life Technologies) with 4′,6-diamidino-2-phenylindole (for nuclear counterstaining). Fluorescent images were captured with an F-view II monochrome camera (Olympus, U-CMAD3) mounted on an Olympus BX51 microscope.
Polylysine d coated coverslips
Manufactured by Merck Group
Sourced in United States
Polylysine D-coated coverslips are laboratory equipment designed for cell culture applications. They provide a specialized surface that promotes cell attachment and growth. The polylysine coating enhances the adhesion of various cell types to the coverslip, facilitating research and experimentation.
Automatically generated - may contain errors
Lab products found in correlation
Antifading prolong gold solution, by Thermo Fisher Scientific (2 mentions)
Bovine serum albumin (bsa), by Merck Group (2 mentions)
Formalin, by Thermo Fisher Scientific (1 mentions)
Cleaved parp, by Thermo Fisher Scientific (1 mentions)
Bx51 microscope, by Olympus (1 mentions)
U cmad3, by Olympus (1 mentions)
Alexa fluor 647, by Thermo Fisher Scientific (1 mentions)
Axiocam mrm camera, by Zeiss (1 mentions)
Observer z1 microscope, by Zeiss (1 mentions)
2 protocols using polylysine d coated coverslips
1
Immunofluorescence Analysis of NF-κB Pathway
2
Quantifying DNA Damage Markers
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3 × 105 cells were adhered onto polylysine D -coated coverslips (Sigma-Aldrich) during a 45-min incubation at 37 °C, fixed in 10% formalin (Fisher Scientific, Pittsburgh, PA, USA) and permeabilized in 1% Triton-X 100 in PBS. Coverslips were blocked for 30 min in 5% bovine serum albumin (Sigma-Aldrich) in PBS with 0.1% Tween-20, probed with γH2AX (S139), phospho-RPA32 (S4/S8) or Ki-67 antibodies (conjugated with AlexaFluor647; Life Technologies) and then with DyLight 488 goat anti-rabbit antibodies (Thermo Scientific, Carlsbad, CA, USA). Coverslips were mounted with anti-fading ProLong Gold Solution (Life Technologies) with 4′,6-diamidino-2-phenylindole (DAPI, for nuclear counterstaining). Fluorescent images were captured with a Zeiss AxioCam MRm camera mounted on a Zeiss Observer.Z1 microscope (Zeiss, Jena, Germany). Cells positive for γH2AX or phospho-RPA were counted in six high-power fields per condition ( × 20 magnification) and referenced to the total number of cells quantified using ImageJ software (National Institutes of Health, Bethesda, MD, USA).
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