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技术文章
Alcian Blue Staining Kit (ABlue)
点击次数：265 发布时间：2019/6/10 8:39:28
Product Description
Alcian Blue is a dye used to detect cell chondrogenesis ａnd it stains the sulfated proteoglycan in cartilage
tissue. The Alcian Blue Staining Kit contains 1% Alcian Blue Solution, 0.1% Nuclear Fast Red in convenient,
ready-to-use solutions. At pH 2.5, Alcian Blue stains acid mucopolysaccharides ａnd shows a blue color, while
Nuclear Fast Red stains nuclei pink to red ａnd the cytoplasm pale pink [1].
Kit Components
Cat. No. # of vials Name Quantity Storage
8378a 1 1% Alcian Blue Solution 100 mL Room temperature
8378b 1 0.1% Nuclear Fast Red 100 mL Room temperature
8378c 1 3% Acetic Acid 100 mL Room temperature
8378d 1 Xylene Substitute 100 mL Room temperature
Materials Supplied by User
Formaldehyde-fixed ａnd paraffin-embedded tissue sections
Ethanol (100%, 95%, 70%, 50%)
Deionized H2O (diH2O)
Product use
ABlue is for research use only. It is not approved for human or animal use, or for application in clinical or in vitro
diagnostic procedures.
Shipping
Room temperature.
References
[1] Steedman, H.F. (1950) Alcian blue 8GS: A new stain for mucin. Quarterly Journal of Microscopic Science,
Vol 91, p477-479.
Procedures
1. Deparaffinize ａnd hydrate slides:
1) Deparaffinize the tissue sections in Xylene Substitute (Cat. #8378d), 3 changes of 5 minutes per change.
2) Hydrate in 100% ethanol, 2 changes of 2 minutes per change.
3) Hydrate in 95% ethanol, 2 changes of 2 minutes per change.
4) Hydrate in 70% ethanol for 2 minutes.
5) Hydrate in 50% ethanol for 2 minutes.
1
6) Rinse in diH2O for 5 minutes.
2. Incubate in 3% Acetic Acid (Cat. #8378c) for 3 minutes.
2. Stain in 1% Alcian Blue Solution, pH 2.5 (Cat. #8378a) for 30-60 minutes.
3. Wash in running tap water for 2 minutes ａnd rinse in diH2O.
4. Stain in 0.1% Nuclear Fast Red (Cat. #8378b) for 10-20 minutes.
5. Wash in running tap water for 1 minute ａnd rinse in diH2O for 2 minutes.
6. Dehydrate ａnd clear slides:
1) Dehydrate in 95% ethanol, 2 changes of 2 minutes per change.
2) Dehydrate in 100% ethanol, 2 changes of 2 minutes per change.
3) Clear the tissue sections in Xylene Substitute (Cat. #8378d), 2 changes of 2 minutes per change.
7. Mount the tissue sections ａnd observe under microscope.
(a) (b)
Figure 1. (a) HDF-f were cultured as pellets in complete MSC Chondrogenic Differentiation Medium
(MCDM, Cat. # 7551) for 50 days. The pellets were fixed in 4% paraformaldehyde ａnd sectioned.
Alcian Blue staining demonstrated the presence of cartilage in cells (Magnification: 10X).
(b) Human Dermal Fibroblasts-fetal (HDF-f, Cat. #2300) were cultured as pellets in growth medium,
complete Fibroblast Medium (FM, Cat. #2301) for 50 days. The pellets were fixed in 4%
paraformaldehyde ａnd sectioned. Alcian Blue staining was not detected (Magnification: 10X).
原创作者：上海雅吉生物科技有限公司
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