Black-Gold® II Stain

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货号：AG400

品牌：Millipore 密理博

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北京朝阳

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商品介绍
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Replacement Information

重要规格表

Key Applications
IHC

Description
Catalogue Number	AG400
Replaces	AG390
Trade Name	BLACK-GOLD
Description	BLACK-GOLD® II Stain
Overview	Black-Gold® II stain is an aurohalophosphate complex which stains specifically for myelin within the central nervous system. Black-Gold II staining provides high resolution, high contrast, a short histochemical processing time, and high reproducibility. Black-Gold II stain is a new and improved version of its predecessor, Black-Gold stain (discontinued). The advantages of Millipore’s Black-Gold II over Black-Gold I stain are that it is more readily soluble, can be utilized at a higher concentration, produces a more uniform and consistent staining, takes less time to stain, and does not require post-staining intensification steps. The use of Black-Gold II stain is tailored to studies using formalin-fixed, non-solvent processed tissue. The technique stains large myelinated tracts dark red-brown, while the individual myelinated fibers appear black. This novel tracer can be used to localize both normal and pathological myelin. Black-Gold II stain can demonstrate and characterize specific myelin changes associated with exposure to diverse neurotoxicants including kainic acid, methamphetamine, acrylamide, domoic acid, 3-nitropropionic acid, Fluoro-Gold™ tracer and isoniazid. Black Gold II Purity: No detectable amount of uncomplexed gold was found. Black Gold II Illumination: Bright field or dark field. Black Gold II Solubility: Freely soluble in water, saline, or dilute acids.

References

Product Information
Components	Black-Gold II: 150mg dry powder, yellow in color.
Presentation	See Components

Applications
Key Applications	Immunohistochemistry
Application Notes	Tissue Fixation: Tissue can be fixed with either formalin or 4% PFA. Typically animals are perfused with neutral phosphate buffered 10% formalin (or 4% formaldehyde) via the ascending aorta. The brains are then removed and fixed overnight in the same fixative solution. Longer fixation times are generally not recommended. For cryosections, the brains may be protected through treatment with sucrose solution (20-30 percent). Subsequently frozen brain tissue is cut on a freezing sliding microtome. Note: Black-Gold II will not stain paraffin embedded or unfixed tissue. Sectioning: Either frozen or fixed, non-frozen vibratome sections can be used and should be cut at a thickness of 20-40 μm. The sections are then typically mounted on positively charged slides. The sections can also be stained loose, although the sections are easier to handle when mounted on slides. Solutions: Black-Gold II powder should be resuspended with saline solution (0.9% NaCl) to a final concentration of 0.3%. Simply add the 150 mg of Black-Gold II powder into 50 mls of 0.9% NaCl solution and mix to resuspend. Once resuspended the Black-Gold II staining solution should be stored in the dark at 4°C and is good for up to 3 months. Staining: 1) Formalin-fixed, non-embedded tissue sections are first mounted from distilled water onto gelatin-coated slides and then air dried at 50-60ºC for at least ½ hour on a slide warmer. 2) Tissue sections are then rehydrated in distilled Milli-Q® water for about 2 minutes. 3) The slides are then transferred to a 0.3% Black-Gold II solution dissolved in 0.9% saline vehicle and heated to 60-65ºC. After about 12 minutes, microscopic monitoring of the extent of the labeling is recommended. This monitoring should be repeated every 2-3 minutes until the desired degree of myelin impregnation is observed (see below). 4) Rinse the slides for about 2 min in distilled Milli-Q water. 5) Transfer the slides to a 1% sodium thiosulfate solution for 3 minutes. 6) Rinse the slides with either three, five min changes of tap water or 15 mins of running tap water. Post-staining steps: 7) Dehydrate sections either via graduated alcoholic solutions or by air drying on a slide warmer. 8) Immerse sections for 1-2 min in xylene or histoclear and then coverslip with a non-polar mounting media.

Applications

Key Applications

Immunohistochemistry

Application Notes

Tissue Fixation:
Tissue can be fixed with either formalin or 4% PFA. Typically animals are perfused with neutral phosphate buffered 10% formalin (or 4% formaldehyde) via the ascending aorta. The brains are then removed and fixed overnight in the same fixative solution. Longer fixation times are generally not recommended. For cryosections, the brains may be protected through treatment with sucrose solution (20-30 percent). Subsequently frozen brain tissue is cut on a freezing sliding microtome. Note: Black-Gold II will not stain paraffin embedded or unfixed tissue.

Sectioning:
Either frozen or fixed, non-frozen vibratome sections can be used and should be cut at a thickness of 20-40 μm. The sections are then typically mounted on positively charged slides. The sections can also be stained loose, although the sections are easier to handle when mounted on slides.

Solutions:
Black-Gold II powder should be resuspended with saline solution (0.9% NaCl) to a final concentration of 0.3%. Simply add the 150 mg of Black-Gold II powder into 50 mls of 0.9% NaCl solution and mix to resuspend. Once resuspended the Black-Gold II staining solution should be stored in the dark at 4°C and is good for up to 3 months.

Staining:
1) Formalin-fixed, non-embedded tissue sections are first mounted from distilled water onto gelatin-coated slides and then air dried at 50-60ºC for at least ½ hour on a slide warmer.
2) Tissue sections are then rehydrated in distilled Milli-Q® water for about 2 minutes.
3) The slides are then transferred to a 0.3% Black-Gold II solution dissolved in 0.9% saline vehicle and heated to 60-65ºC. After about 12 minutes, microscopic monitoring of the extent of the labeling is recommended. This monitoring should be repeated every 2-3 minutes until the desired degree of myelin impregnation is observed (see below).
4) Rinse the slides for about 2 min in distilled Milli-Q water.
5) Transfer the slides to a 1% sodium thiosulfate solution for 3 minutes.
6) Rinse the slides with either three, five min changes of tap water or 15 mins of running tap water.

Post-staining steps:
7) Dehydrate sections either via graduated alcoholic solutions or by air drying on a slide warmer.
8) Immerse sections for 1-2 min in xylene or histoclear and then coverslip with a non-polar mounting media.

Biological Information

Physicochemical Information

Dimensions

Materials Information

Toxicological Information

Safety Information according to GHS

Safety Information

Product Usage Statements
Usage Statement	Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.

Storage and Shipping Information
Storage Conditions	The kit can be stored at room temperature. Ideally, the Black-Gold II powder should be kept in a desiccator, because of its hygroscopic nature. The dry powder, if unopened and stored properly, is usable for up to one year from date of receipt. Once resuspended the Black-Gold II staining solution should be stored in the dark at 4°C and is good for up to 3 months. TOXICITY: Although the compound appears to be of low toxicity, it has not been extensively evaluated and therefore routine laboratory caution should be exercised. Not intended for human consumption.

Storage and Shipping Information

Storage Conditions

The kit can be stored at room temperature. Ideally, the Black-Gold II powder should be kept in a desiccator, because of its hygroscopic nature. The dry powder, if unopened and stored properly, is usable for up to one year from date of receipt. Once resuspended the Black-Gold II staining solution should be stored in the dark at 4°C and is good for up to 3 months.

TOXICITY: Although the compound appears to be of low toxicity, it has not been extensively evaluated and therefore routine laboratory caution should be exercised. Not intended for human consumption.