Immunofluorescence (IF) staining was performed following our published conditions [64 (link),65 (link),66 (link)]. Briefly, cells were fixed with prechilled (−20 °C) acetone–methanol for 15 min prior to the addition of primary antibodies anti-γH2AX (1:100, Cell Signaling, Danvers, MA, USA) and anti-phospho-ATM (S1981) (1:100, Cell Signaling) at 4 °C overnight. After rinsing, FITC-Donkey anti-mouse IgG (1:200, Jackson Immuno Research Lab, West Grove, PA, USA) and Rhodamine-Donkey anti-rabbit IgG (1:200, Jackson Immuno Research Lab) were added for 1 h at room temperature. Slides were mounted using the VECTASHIELD mounting medium containing DAPI (VECTOR Lab Inc., Burlington, ON, Canada). VE-281 (Selleckchem, Burlington, ON, Canada) or KU-55933 (Selleckchem) was added to cells 8 h prior to treatment. Images were then acquired with a fluorescent microscope (Axiovert 200, Carl Zeiss, North York, ON, Canada).
Rhodamine donkey anti rabbit igg
Manufactured by Jackson ImmunoResearch
Sourced in United States
Rhodamine-Donkey anti-rabbit IgG is a secondary antibody conjugated with the fluorescent dye rhodamine. It is designed to bind to and detect rabbit primary antibodies in various immunoassays and imaging applications.
Automatically generated - may contain errors
Lab products found in correlation
Axiovert 200, by Zeiss (2 mentions)
Fitc donkey anti mouse igg, by Jackson ImmunoResearch (1 mentions)
Ku55933, by Selleck Chemicals (1 mentions)
Anti phospho atm s1981, by Cell Signaling Technology (1 mentions)
Anti γh2ax, by Cell Signaling Technology (1 mentions)
Polyclonal anti flag, by Merck Group (1 mentions)
2 protocols using rhodamine donkey anti rabbit igg
1
Immunofluorescence Staining for DNA Damage
2
Immunofluorescence Staining of Epitope-Tagged Proteins
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Immunofluorescence (IF) staining was performed by fixing cells with paraformaldehyde for 20 min, followed by permeabilization with 0.3% Triton X-100 for 15 min. Cells were then blocked with a blocking buffer (3% donkey serum + 3% bovine serum albumin (BSA) + 0.3% Triton X-100 in PBS) prior to the addition of primary antibodies polyclonal anti-FLAG (a commonly used polypeptide epitope tag) (1:100, Sigma) and monoclonal anti-HA (a tag epitope derived from hemagglutinin) (12CA5, 1:10 of hybridoma conditional medium) at 4°C overnight. After rinsing with PBS, secondary antibodies FITC (fluorescein)-donkey anti-mouse immunoglobulin (Ig)G (1:200, Jackson Immuno Research Lab) and rhodamine-donkey anti-rabbit IgG (1:200, Jackson Immuno Research Lab) were applied for 1 h at room temperature. Slides were eventually mounted using VECTASHIELD mounting medium containing 4′,6-diamidino-2-phenylindole (DAPI; VECTOR Lab Inc.). Images were taken with a fluorescent microscope (Axiovert 200, Carl Zeiss).
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