ArticleHistochemistry and cell biology2011
Antigen retrieval to improve the immunocytochemistry detection of sigma-1 receptors and ER chaperones.
Article in Histochemistry and cell biology, 2011. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 11 papers.
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Who cites it
11 citing papers in PubMed, 23 citations in OpenAlex.
- Sigma-1 Expression in Chronic Mouse Models of Temporal Lobe Epilepsy.Molecular neurobiology · 2026Article
- Mitochondrial‑endoplasmic reticulum crosstalk: Molecular mechanisms and implications for cardiovascular disease (Review).Molecular medicine reports · 2025Review
- Actin Filament in the First Cell Cycle Contributes to the Determination of the Anteroposterior Axis in Ascidian Development.Journal of developmental biology · 2022Article
- Dynamic changes in the association between maternal mRNAs and endoplasmic reticulum during ascidian early embryogenesis.Development genes and evolution · 2022Article
- Impaired SorLA maturation and trafficking as a new mechanism for SORL1 missense variants in Alzheimer disease.Acta neuropathologica communications · 2021Article
- Wildtype σ1 receptor and the receptor agonist improve ALS-associated mutation-induced insolubility and toxicity.The Journal of biological chemistry · 2020Article
- Antigen retrieval pre-treatment causes a different expression pattern of Cav3.2 in rat and mouse spinal dorsal horn.European journal of histochemistry : EJH · 2019Article
- Mitochondria-associated membrane collapse is a common pathomechanism in SIGMAR1- and SOD1-linked ALS.EMBO molecular medicine · 2016Article
- Altered sigma-1 receptor expression in two animal models of cognitive impairment.Molecular imaging and biology · 2015Article
- Sigma-1 receptor expression in sensory neurons and the effect of painful peripheral nerve injury.Molecular pain · 2013Article
- Recent progress in histochemistry and cell biology.Histochemistry and cell biology · 2012Review
Corrections and comments
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Authors and funding
5 authors at 2 institutions in 1 country.
Funding
Abstract
Molecular chaperones localized at the endoplasmic reticulum (ER) lumen constitutively or cellular stress-dependently associate with a variety of proteins to promote their proper folding or to inhibit protein misfolding. ER chaperones preferentially form large complexes with co-chaperones and/or misfolded proteins in a highly crowded cellular environment that often hampers their detection by immunocytochemistry (ICC). This study establishes an antigen retrieval (AR) protocol to improve the ICC detection of ER chaperones in cultured cells using widely available antibodies against synthetic peptides. Among ten different antigen retrieval/fixation conditions, only the AR with Tris-HCl (pH 9.5) containing 6 M urea (80°C for 10 min) significantly improved the ICC detection of the novel ER chaperone sigma-1 receptor (Sig-1R) in Chinese hamster ovary cells. Extended fixation with 4% paraformaldehyde for 1 h effectively preserved the morphology of the ER under the AR condition. This method greatly enhanced the signal-to-noise ratio in Sig-1R ICC, thus allowing for semi-quantitative detection of protein upregulation under ER stress. The AR similarly improved the ICC detection of a series of other major ER chaperones, including BiP/GRP78, GRP94, calnexin, calreticulin, ERp57, protein disulfide isomerase, and cyclophilin B. The improved ICC methodology using the urea AR at 80°C may improve ICC of ER molecules as well as visualization of ER structure and substructures.
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Registered trials
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