Related Experiment Video
Updated: Jun 5, 2026
Investigating Mast Cell Secretory Granules; from Biosynthesis to Exocytosis
Published on: January 26, 2015
Effects of endogenous substance P expression on degranulation in RBL-2H3 cells
Hua Hu1, Ruxin Zhang, Xiaoyun Fang
1Department of ENT, Huadong Hospital, Shanghai Medical School, Fudan University, Shanghai, 200032, People's Republic of China.
Objective And Design:
To determine whether RBL-2H3 cells have endogenous substance P (SP) expression under immunoglobulin E (IgE)-activated and inactivated conditions, and to ascertain the function of endogenous SP in the antigen-induced degranulation of RBL-2H3 cells.
Materials And Methods:
SP mRNA and protein expression in both inactivated and 2,4-dinitrophenol (DNP)-specific IgE-activated RBL-2H3 cells were assessed by real-time PCR and immunofluorescence, respectively. Following activation with DNP-specific IgE, the degranulation of RBL-2H3 cells in response to DNP-bovine serum albumin (BSA), with and without endogenous SP expression, was assessed by monitoring the release of the granular enzyme β-hexosaminidase.
Results:
Endogenous SP mRNA and peptide expression increased in activated RBL-2H3 cells, compared with inactivated RBL-2H3 cells. The small hairpin RNA (shRNA)-mediated knockdown of endogenous SP reduced the degranulation ability of RBL-2H3 cells.
Conclusions:
Activated RBL-2H3 cells express endogenous SP which plays a role in antigen-induced degranulation.
More Related Videos
11:31Isolation of Peritoneum-derived Mast Cells and Their Functional Characterization with Ca2+-imaging and Degranulation Assays
Published on: July 4, 2018
17:35A Microplate Assay to Assess Chemical Effects on RBL-2H3 Mast Cell Degranulation: Effects of Triclosan without Use of an Organic Solvent
Published on: November 1, 2013