EDTA Decalcification Solution (IHC)
MCE EDTA Decalcification Solution (IHC) consists primarily of disodium EDTA and phosphates and has a pH of 7.2. The product uses chelation-based decalcification at neutral pH; compared with acid-based decalcifying solutions, it has less impact on tissue structure and antigenicity, and is therefore suitable for subsequent immunohistochemical (IHC) staining and in situ hybridization. Note that decalcification with EDTA is relatively slow—the time required depends on tissue type, specimen size, and degree of calcification—and it is therefore not suitable for experiments that require rapid decalcification.
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Storage :Room temperature, 1 year.
- Manual (2167KB)
- COA
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Description & Advantages
EDTA Decalcification Solution is a mild decalcifying reagent suitable for the decalcification of bone, teeth, and other calcified tissues, preparing them for subsequent tissue embedding and sectioning. EDTA forms soluble chelate complexes with calcium ions in the tissue and gradually removes calcium salts, thereby achieving decalcification while preserving tissue morphology, cellular structure, and antigenicity as much as possible.
MCE EDTA Decalcification Solution (IHC) consists primarily of disodium EDTA and phosphates and has a pH of 7.2. The product uses chelation-based decalcification at neutral pH; compared with acid-based decalcifying solutions, it has less impact on tissue structure and antigenicity, and is therefore suitable for subsequent immunohistochemical (IHC) staining and in situ hybridization. Note that decalcification with EDTA is relatively slow—the time required depends on tissue type, specimen size, and degree of calcification—and it is therefore not suitable for experiments that require rapid decalcification.
Protocols
1. Tissue Sampling: Prepare bone specimens approximately 5 mm thick. Avoid excessively thick specimens.
2. Washing after Fixation: Following fixation, wash the tissue 3 times with PBS for 20 min each.
3. Distilled Water Washing: Wash the tissue 3 times with distilled water for 20 min each.
4. Decalcification: Immerse the tissue in 20–30 tissue volumes of EDTA Decalcification Solution. Replace the solution weekly until the decalcification endpoint is reached. Decalcification typically requires 10–30 days or longer; some specimens may require 2 weeks to 3 months, depending on tissue thickness and degree of calcification.
5. Assessment of the Decalcification Endpoint: Assess tissue softening by gentle needle probing, light finger pressure, or gentle compression with forceps. Decalcification may be stopped when the bone tissue has softened sufficiently or a needle penetrates without appreciable resistance. Perform these checks gently and avoid excessive force or repeated testing to minimize damage to tissue architecture.
6. Washing after Decalcification: Once decalcification is complete, rinse the tissue thoroughly several times with distilled water.
7. Dehydration and Embedding: Dehydrate and embed the tissue according to standard histological procedures.
Note: To accelerate decalcification, incubate at 37°C. Alternatively, the following microwave-assisted decalcification conditions may be used as a reference: approximately 200 W for 5 min per heating cycle, with an interval of 3–5 min between cycles. Adjust the number of heating cycles according to tissue thickness and density.
Storage
Room temperature, 1 year.
Attention
1. The decalcification time depends on tissue thickness and degree of calcification. Check progress regularly to avoid incomplete or excessive decalcification.
2. Moderate warming accelerates decalcification. The temperature should generally not exceed 37-40°C and must not exceed 60°C to avoid tissue damage.
3. Use glass containers and avoid metal containers.
4. Fix tissues before decalcification or perform fixation and decalcification simultaneously. Avoid fixation only after decalcification.
5. This product is for R&D use only, not for drug, household, or other uses.
6. For your safety and health, please wear a lab coat and disposable gloves to operate.
Components
| Components | HY-K2011-500 mL | HY-K2011-1 L | HY-K2011-3 L | |
|---|---|---|---|---|
| EDTA Decalcification Solution (IHC) | 500 mL | 1 L | 3 L |
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