Plant Protein and DNA/RNA Co-Localization (IF-FISH) Service

CD BioSciences enables co-localization of nucleic acid and plant proteins in cells by fluorescence in situ hybridization (FISH) and immunofluorescence (IF) double staining, characterizing the presence of nucleic acid-protein interactions from a co-localization perspective. Our technology balances the conflicting requirements for permeability, fixation, and antigenic preservation to detect nucleic acid and protein expression with high resolution and sensitivity.

FISH utilizes the immunochemical reaction between reporter molecules and fluorescein-labeled specific affinities or directly via a fluorescent detection system for qualitative, semi-quantitative, or relative localization of nucleic acids (mRNA, lncRNA, circRNA, miRNA).

IF labels antibodies with fluorescent substances for antigen localization. It is used to detect or localize various antigens and can also bind to proteins for the localization of antibodies.

  Figure 1. Workflow diagram for ISH, FISH, and IF/FISH. (Zimmerman, S. G., et al., 2013)

Service Content

Workflow

• Fixed tissues, dehydrated, and sectioned, paraffin sections deparaffinized and hydrated.
• Boil-treated sections are digested dropwise with proteinase K.
• Pre-hybridization solution incubation.
• Hybridization with probe-containing hybridization solution.
• SCC wash after hybridization.
• Incubate primary and secondary antibodies.
• Using DAPI to re-stain the nucleus.
• Microscopic examination for photographs.

Fluorescent dyes we offer

IF-FISH optimization services

• Sample preparation optimization.

We screened the appropriate stimulation time during specimen preparation and used colchicine stimulation to obtain the intermediate chromosome division image.

• Selection of proteinase K concentration.

We provide the appropriate concentration of proteinase K action. A high concentration affects the morphology of the nucleus, while a low concentration makes it difficult for the probe to enter the nucleus and reduces the hybridization rate.

• Optimization of hybridization conditions.

We select appropriate hybridization temperature, time, and incubation time for clients.

• Temperature control of rinsing solution.

The FISH experiment is a very temperature-sensitive experiment. We ensure the optimal temperature of the rinsing solution during our experiments.

Sample Requirements

CD BioSciences is committed to providing cutting-edge IF-FISH services to clients. We offer superior solutions to visualize and characterize the localization and interaction of proteins and nucleic acids in plant cells. If you have requirements in studying plant protein interactions, please contact us. Our team of highly skilled scientists will provide you with precise and detailed results.

For research use only, not for clinical use.