General Information:

Grade: Molecular Biology
Contents: Formamide, SSC, Denhardt’s, Dextran Sulfate, SDS, Yeast tRNA and Salmon sperm DNA in distilled water
Not hazardous product

Product Description:

Ready-to-use hybridization solution optimized for in situ hybridization (ISH) on brain and spinal cord sections. The hybridization solution consists of 50% formamide, 5× SSC, 5× Denhardt’s solution, 10% dextran sulfate, 0.5% SDS, 100 µg/ml yeast tRNA and 100 µg/ml salmon sperm DNA. The solution can be used effectively with either non-radioactively or radioactively labeled RNA, oligonucleotide, and DNA probes.

Product Features:  

• Compositions: 50% formamide, 5× SSC, 5× Denhardt’s solution, 10% dextran sulfate, 0.5% SDS, 100 µg/ml yeast tRNA and 100 µg/ml salmon sperm DNA
• Formulated from molecular grade reagents. Made with 0.1 micron sterile filtered distilled water
• Buffer solution are free of DNase, RNase, and protease activity
• Optimized for in situ hybridization on brain and spinal cord sections
• Increase the sensitivity of hybridization. Reduces the time of hybridization
• Ready-to-use, store at freezer (-20°C to -80°C)
• HDPE bottle (temperature -100°C to 120°C)
• For Research Use Only

Approved Results:

Demonstration of the utility of Hybridization Solution (Cat # 007611) for in situ hybridization (ISH) and ISH combined with immunohistochemistry (IHC) on brain sections fixed with 4% PFA.   (A) GAD mRNA-expressing cells in the hippocampus. (B) Section was counterstained with Neutral Red (Cat # 003034). Arrowhead points to a mRNA-expressing cell. (C) 4% PFA (Cat # 007512)-fixed sections (20 micron) were first subjected to ISH, followed by IHC. Arrows in C1 denote dual-labeled cells. One arrow-pointed cell was magnified in C2. In the ISH procedures, 20× SSC (Cat # 007603), 10× TBS (pH7.0)(Cat # 007508), 1× TBS (pH 9.5) (Cat # 007510), and distilled water (Cat # 007615) from Bioenno Tech were employed. Standard avidin-biotin-peroxidase IHC was performed on free-floating sections. Bioenno DAB-Cobalt Substrate Kit (Cat # 003843) was used in the development of immunoreactive signals.

Instructions for Use (4% PFA fixed brain sections as an example):

• Collection of sections: collect sections in DEPC-treated 2× SSC at RT (15-25°C);
• Pre-hybridization: (a) transfer the sections to solution composed of 2× SSC/Hybridization Solution (1:1) and incubate for 30-60 min at RT; (b) transfer the sections to Hybridization Solution and incubate for 1 hour at 50-70°C;
• Hybridization: transfer the sections to Hybridization Solution with DIG- or biotin-labeled probe. Incubate the sections at 50-70°C for 8-24 hours.

Notes: The incubation and hybridization can be performed in sterile six-well plates. Please use RNAse-free solutions. The temperature and time of incubations should be determined and optimized by the investigator.

• Wash of sections and detection of signals: wash the sections in 2× SSC, 50% formamide/2× SSC, 50% formamide/0.1× SSC, and 0.1× SSC at room temperature or 60-70°C; Hybrid molecules can be detected with anti-DIG or anti-biotin serum.

Shipping, Storage, Safety, and Handling Precautions:

• Shipped at room temperature. Store the solution in freezer (-20 to -80°C).
• The solution is designed for in vitro research use only and not for drug, diagnostic or other uses.
• The solution contains reagents that may be harmful in contact with skin, by inhalation or ingestion. Do not pipette by mouth. Use ordinary precautions to avoid inhalation and contact with skin and eyes. In case of contact, wash immediately with generous amounts of water and seek medical advice. If swallowed, wash out mouth with water and immediately call a physician.
• Perform experiment under a chemical hood. Wear suitable protective clothing, gloves and eye/face protection. Wash hands thoroughly after performing the experiment.
• Material safety data sheet (MSDS) is available upon request.

Warranty: 12 months from the date of purchase.