贴壁细胞免疫荧光(间接染色) 方法

Ran Chen

doi:10.21769/BioProtoc.50

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Peer-reviewed

Immunofluorescence (Indirect Staining) Protocol for Adherent Cells

贴壁细胞免疫荧光(间接染色) 方法

RC Ran Chen email

发布: 2012年02月05日第2卷第3期 DOI: 10.21769/BioProtoc.50 浏览次数: 25471

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Abstract

In this protocol, different types of adherent cells are fixed to coverslips by 4% paraformaldehyde. Target proteins are then stained by specific antibodies and fluorescent 2^nd antibodies. This protocol therefore provides a method for immunostaining of adherent cells.

Keywords: Immunofluorescence (免疫荧光)

Cell (细胞)

Adherent (贴壁)

Materials and Reagents

Raw264.7, MCF-7 or Hela cells
DPBS (Life Technologies, Invitrogen^TM, catalog number: 14190-250 )
FBS (Atlanta Biologicals, catalog number: S11110H )
Anti-fade mounting medium: e.g. ProLong Gold Antifade Mountant (Life Technologies, Invitrogen^TM, catalog number: P10144 ) with DAPI (if nuclear staining is needed) or without DAPI
Paraformaldehyde (Sigma-Aldrich, catalog number: 158127 )
General chemicals (Sigma-Aldrich)
Triton X-100 (Sigma-Aldrich, catalog number: T8787 )
Tween 20 (Sigma-Aldrich, catalog number: P2287 )
4% paraformaldehyde-freshly-prepared (5 ml) (see Recipes)
Antibodies (see Recipes)