Anti-freeze Cryoprotectant Solution for Fixed Sections

Laboratory Use Only

Product Package:

1) 250 ml per unit (Catalog No.: 006799-250)

2) 1,000 ml per unit (Catalog No.: 006799-1L)

3) 1 Gallon per unit (Catalog No.: 006799-1G)

General Information:

• Grade: Biology
• Contents: Ethylene glycol and Glycerol in phosphate buffer
• Not hazardous product

 Product Description:

Ethylene glycol based cryoprotectant prepared in phosphate buffer (PB, pH 7.4). This solution is optimal for long-term storage of brain and spinal cord sections fixed with aldehyde-based fixatives, e.g., 4% PFA (paraformaldehyde), formalin, and glutaraldehyde. Transfer 4% PFA-fixed and unstained sections to the cryoprotectant and store at -20°C for 2 years, which produces no adverse effects on tissue morphology, and most of fluorescent proteins and antigen epitopes. Stored sections can be subjected for regular histology, immunofluorescence and immunohistochemistry (IF and IHC).

Product Features: 

• Cryopreservation of brain and spinal cord sections
• High-purity ethylene glycol (30%) and glycerol (20%) in 0.02 M PB at pH 7.4
• No DMSO (dimethyl sulfoxide), no sucrose, and no formaldehyde
• Optimized for histology, IF, and IHC
• Ready-to-use, the solution itself can be stored at -20°C to 25°C
• HDPE bottle (temperature -100°C to 120°C)
• For Research Use Only

 Approved Results: 

Demonstration of the utility of anti-freeze cryoprotectant (Cat # 006799) for immunohistochemistry (IHC) and immunofluorescence staining (IF) on brain sections fixed with 4% PFA (Cat # 006775).

(A1) Freshly harvested section was stained with a rabbit anti-Fos from Oncogene Sciences. (A2) An adjacent section seated in the cryoprotectant for 2 years (-20°C) was stained using the same antibody. (B, C) Examples of dual-labeling on adult rat (B) and mouse (C) brain sections that were stored in the Cryoprotectant for 2 years.

Standard avidin-biotin-peroxidase IHC (A, B) and convenient IF (C) were performed on free-floating sections. Bioenno DAB-Cobalt Substrate Kit (Cat # 003843) was used together with BDHC Kit (Cat # 003850) to present single- and/or dual-labelling.

Instructions for Use:

• Fixed tissue block can be sectioned at 10-40 μm thickness using a cryostat.
• The sections can be

(a) first collected in 0.1 M PB (pH 7.4), and then transfer to the cryoprotectant; or

(b) directly collected in the cryoprotectant.

• Seal or cove tightly the cryoprotectant with sections, and store at -20°C.

Shipping, Storage, Safety, and Handling Precautions:

• Shipped at room temperature.
• Store the solution at -20°C – 25°C and avoid strong direct light. The shelf life at 25°C is one year.
• 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.

Return Policy: Bioenno Tech’s return policy for this product is 90 days from the date of purchase.

Free Technical Support: Email your questions to contact@bioenno.com

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