IMMUNO SHOT immunostaining

Protocol for cultured cells


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An example of cell staining using fluorescent-labeled secondary antibody is described.

1) Remove the culture medium and wash cultured cells once with PBS.

2) Fix cells for 30 min with 4% paraformaldehyde in 0.1 M phosphate buffer at RT.

3) Wash cells with PBS for 5 min three times.

4) Add blocking solution, and incubate for 30 min at RT.

5) Wash cells with PBS for 5 min three times.

6) Dilute primary antibody by one of three solutions of IMMUNO SHOT Immunostaining to the concentration recommended by supplier., and incubate for 1 hours at RT or overnight at 4C in humid chamber.

7) Wash cells with PBS for 5 min three times.

8) Dilute fluorescent dye–labeled secondary antibody by one of three solutions of IMMUNO SHOT Immunostaining to the concentration recommended by supplier, and incubate for 1 hours at RT.

9) Wash cells with PBS for 5 min three times.

10) Observe the cells by fluorescent microscope.

(Caution: the addition of serum or protein into IMMUNO SHOT Immunostaining alters the nature of the reagent, and affects the efficacy of the reagent)

IMMUNO SHOT immunotaining top

To be used for research only. DO NOT use for human gene therapy or clinical diagnosis.


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