Optimized immunofluorescence staining protocol to detect the nucleoporin Nup98 in different subcellular compartments

2009 ◽  
Author(s):  
Mohamed Kodiha ◽  
Rehan Umar ◽  
Ursula Stochaj
Keyword(s):  
Immunofluorescence Staining ◽  
Subcellular Compartments ◽  
Staining Protocol

scholarly journals cFos immunoreactivity is enhanced with biotin amplification.

1994 ◽  
Vol 42 (12) ◽  
pp. 1635-1642 ◽  
Author(s):  
K A Berghorn ◽  
J H Bonnett ◽  
G E Hoffman
Keyword(s):  
Signal To Noise Ratio ◽  
Immediate Early Gene ◽  
Immunofluorescence Staining ◽  
Early Gene ◽  
Primary Antibody ◽  
Immediate Early ◽  
High Signal ◽  
Fluorescence Staining ◽  
Conventional Procedure ◽  
Staining Protocol

Through modification of the protocol by Adams, we developed a biotin amplification procedure for immunofluorescence staining of immediate early gene proteins and also applied biotin amplification for metal enhancement of diaminobenzidine staining in an immunoperoxidase protocol. Commercially available anti-cFos antisera were used to compare conventional "Elite" avidin-biotin complex reactions with biotin amplification reactions (visualized with peroxidase staining or streptavidin-Texas Red fluorescence). Biotin amplification and peroxidase staining (with or without nickel salts) enabled detection of cFos in stimulated neurons with primary antibody concentrations five- to tenfold lower than the conventional procedure. With immunofluorescence staining, at primary antibody concentrations too low to detect cFos with the conventional biotin-streptavidin fluorescence staining protocol, biotin amplification enabled clear cFos fluorescence staining with both antisera. The fluorescence staining exhibited a high signal-to-noise ratio and enabled antibody concentrations four times lower than those used for conventional ABC "Elite" peroxidase procedures. In conclusion, the application of biotin amplification to cFos immunocytochemical localization has the promise of aiding the scientist in detecting these immediate early gene products.

scholarly journals Optimized immunofluorescence staining protocol for imaging germinal centers in secondary lymphoid tissues of vaccinated mice

2021 ◽  
Vol 2 (3) ◽  
pp. 100499
Author(s):  
Sigrid Fra-Bido ◽  
Simon A. Walker ◽  
Silvia Innocentin ◽  
Michelle A. Linterman
Keyword(s):  
Germinal Centers ◽  
Immunofluorescence Staining ◽  
Lymphoid Tissues ◽  
Staining Protocol

scholarly journals Immunofluorescence staining protocol for STED nanoscopy of Plasmodium-infected red blood cells

2018 ◽  
Author(s):  
Ann-Kathrin Mehnert ◽  
Caroline Sophie Simon ◽  
Julien Guizetti
Keyword(s):  
Red Blood Cells ◽  
Blood Cells ◽  
Single Cells ◽  
Blood Stage ◽  
Immunofluorescence Staining ◽  
Cell Seeding ◽  
Cell Integrity ◽  
Nuclear Pores ◽  
Staining Protocol ◽  
Intranuclear Microtubules

AbstractImmunofluorescence staining is the key technique for visualizing organization of endogenous cellular structures in single cells. Labeling and imaging of blood stage Plasmodium falciparum has always been challenging since it is a small intracellular parasite. The gold standard for parasite immunofluorescence is fixation in suspension with addition of minute amounts of glutaraldehyde to the paraformaldehyde-based solution. While this maintains red blood cell integrity, it has been postulated that antigenicity of the parasite proteins was, if at all, only slightly reduced. Here we show the deleterious effect that even these small quantities of glutaraldehyde can have on immunofluorescence staining quality and present an alternative cell seeding protocol that allows fixation with only paraformaldehyde. The highly improved signal intensity and staining efficiency enabled us to carry out RescueSTED nanoscopy on microtubules and nuclear pores and describe their organization in greater detail throughout the blood stage cycle.Highlights- Omitting glutaraldehyde from fixative improves immunofluorescence staining- STED nanoscopy is readily applicable to infected red blood cells- Intranuclear microtubules are nucleated from distinct sites

scholarly journals TRIPLE (Insulin, Glucagon and EGFP) Immunofluorescence Staining Protocol in Pancreas

BIO-PROTOCOL ◽  
2014 ◽  
Vol 4 (5) ◽  
Author(s):  
Woogyun Choi ◽  
Randal Kaufman ◽  
Sung Back
Keyword(s):  
Immunofluorescence Staining ◽  
Staining Protocol
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