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General Questions
Four-color IHC staining protocol
The protocol described below is a sequential staining process and may be adapted to any desired number of colors by adding…
Tue, Jul 18, 2023
Glossary
Antibody: An immunoglobulin that recognizes some portion of an antigen molecule. The antibody contains a species-specific Fc region and antigen-specific…
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
EGS (ethyleneglycol-bis-succinimidyl-succinate) Fixation
EGS (ethyleneglycol-bis-succinimidyl-succinate) fixation (method to preserve microtubules and membrane bound antigens) Rinse cells with pre-warmed (37°C) PBS. Prepare EGS stock…
Tue, Jul 18, 2023
Formaldehyde Fixation
Formaldehyde fixation (for membrane associated components) Fixative Preparation Dissolve 16 g paraformaldehyde in 80 ml dH2O by stirring at 70°C…
Tue, Jul 18, 2023
Methanol Fixation
Methanol fixation (for cytoskeletal components) The methanol fixation is an easy method. However, it frequently solubilizes and removes membrane bound…
Tue, Jul 18, 2023
Checking for Interference or Linearity
Interference is when some component of a sample prevents the appropriate binding events to take place. Percent recovery is the…
Tue, Jul 18, 2023