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Blue nuclear counterstain signal is dramatically reduced in treated cells.
If the blue nuclear signal decreases by more than 30%, the testing compound is generally cytotoxic. Optimize the concentration of…
Tue, Jul 18, 2023
After incubation with the dual detection reagent at 37° C for 30 min, the cells were not healthy and rounding up. Some cells were also detached from the plate. No specific staining on positive control was observed vs. DMSO controls.
For fragile cells, the following modification to the standard protocol are suggested: Use PBS/5% FBS as an assay buffer or…
Tue, Jul 18, 2023
Intracellular Staining Protocol: Two Step Fix/Perm
Two Step Fixation and Permeabilization Materials: Maintain Reagents and buffers at 4°C. Fluorochrome-conjugated primary antibody Fluorochrome-conjugated isotype control antibody Fixation…
Tue, Jul 18, 2023
Intracellular Staining Protocol: One Step Fix/Perm
One Step Fixation and Permeabilization Materials: Maintain Reagents and buffers at 4°C. Fluorochrome-conjugated primary antibody Fluorochrome-conjugated isotype control antibody Fixation/Permeabilization…
Tue, Jul 18, 2023
Extracellular Staining Protocol
Materials: Maintain Reagents and buffers at 4°C. Fluorochrome-conjugated primary antibody Fluorochrome-conjugated isotype control antibody Fixation Buffer, ADI-950-011 PBS (Phosphate Buffered…
Tue, Jul 18, 2023
Fixation of the Cytoskeleton
Fixation of the cytoskeleton If components of the cytoskeleton are of interest, cells can be extracted prior to fixation. Rinse…
Tue, Jul 18, 2023
Is it possible to detect autophagy by fluorescence microscopy using 3D culture systems?
We have not tested the dye in 3D culture systems, but the small cell clusters are probably dye-accessible. For live…
Tue, Jul 18, 2023
Can any culture medium be used to measure cAMP/cGMP in culture supernatants with cAMP/cGMP ELISA and cAMP/cGMP Complete ELISA kits?
As indicated in the insert, as long as the standard curve is titrated in the same culture media that the…
Tue, Jul 18, 2023
Is the acetylation step really necessary prior to cAMP/cGMP quantification?
Samples should be acetylated if they contain a very low concentration of cAMP or cGMP, or when greater sensitivity is…
Tue, Jul 18, 2023
Paraformaldehyde/Glutaraldehyde Fixation
Paraformaldehyde/glutaraldehyde fixation(method of choice for double labeling of membrane bound and cytoskeletal antigens) Rinse cells with PBS at room temperature.…
Tue, Jul 18, 2023