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G1221-5ML Tissue Autofluorescence Quencher Tissue Autofluorescence Quencher

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G1221-5ML Tissue Autofluorescence Quencher Tissue Autofluorescence Quencher

Immunofluorescence technology uses fluorescent molecule-labeled antibodies to locate the bound antigens, and realize subcellular visualization of proteins, glycans, small biomolecules, and non-biological molecules. During this process, autofluorescence in the tissue tends to severely interfere with observations. Autofluorescence is a general term for background fluorescence signals unrelated to the target signal generated in the process of immunofluorescence detection. , red blood cells, lipofuscin, etc. will produce strong autofluorescence, which significantly affects the signal-to-noise ratio of immunofluorescence experiments. In addition, substances such as formalin and paraformaldehyde used in tissue fixation also produce a large amount of autofluorescence.

 

This product can effectively reduce tissue autofluorescence and improve the signal-to-noise ratio of immunofluorescence detection. The active ingredient of liquid A is copper ion, which can effectively reduce the autofluorescence of red blood cells, and the active ingredient of liquid B is Sudan black, which can significantly reduce the autofluorescence caused by lipofuscin.

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Technical Specification

Storage and transportation

Store at room temperature away from light, valid for 12 months.

 

Instructions

  1. Quenching of autofluorescence before immunolabeling: After antigen retrieval of tissue sections, circle the tissue with histochemical strokes, then dropwise add tissue autofluorescence quencher solution A to cover the tissue, incubate at room temperature for 30 minutes, and wash with pure water for 5 minutes .
  2. The tissue sections are subjected to routine immunofluorescence detection steps such as blocking, primary antibody incubation, secondary antibody incubation, and DAPI counterstaining of cell nuclei.
  3.  Quenching of autofluorescence after immunolabeling: Add solution B of tissue autofluorescence quencher dropwise to cover the tissue, incubate in the dark at room temperature for 5 min, and rinse with running water for 3 min.
  4. Cover slides: wash the slides in PBS with shaking (can be placed on a destaining shaker) 3 times, 5 min each time. The sections were dried a little and then mounted for microscopic examination.

 

 

Precautions

  1. Different samples have different autofluorescence principles, and the effect of using tissue autofluorescence quenchers may vary.
  2. Theoretically, quenchers against autofluorescence will also reduce the fluorescence intensity of antibodies to a certain extent. After testing, the quenching degree of this reagent on autofluorescence far exceeds the reduction of antibody fluorescence intensity, and there is a good balance between the two.
  3. After adding the tissue autofluorescence quencher B solution, the tissue will be stained dark blue, but it will not affect the fluorescence photography.
  4. After incubation in solution B, the slices can be washed with water or PBS, but not with a solution containing Tween, otherwise the solution B will be washed away excessively, which will affect the quenching effect of background fluorescence.
  5. Be sure to tighten the cap of the tissue autofluorescence quencher solution B after each use to prevent the solvent from volatilizing.
  6. Please wear lab coat and disposable gloves during operation.
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