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  Proteinase K Unmasking Solution (10X) 60ml- Available May 1, 2011 Cat# AB10

Catalog # AB10
 
Epitope Unmasking Solution (10X) 500mls

Our Price: $175.00


Product Code: AB10
Qty:

Description Technical Information
 
When heat is not acceptable for your epitope unmasking during your IHC and IF,  ProHisto offers the 10X Proteinase K Unmasking Solution. This solution reverts cross links formed during fixation, and restores epitope configuration, all at room temperature. The Proteinase K Unmasking Solution will enhance the binding of your primary antibody, and result in a strong signal with little background. Can process 200 slides.
Features

  • Epitope Unmasking Solution™ (10X)

  • Catalog number AA5, AA6

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  • Reagent provided

  • A liquid buffer composed of 10% or less of citric acid anhydrous.

  • 250 or  500ml bottle.

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  • Application

  • For  In Vitro diagnostic use.

  • Antigen Retrieval is used to recover antigenicity in formalin-fixed, paraffin-embedded tissues, producing greatly improved staining, and preventing false negative staining of over-fixed tissue. Formulin fixation of tissues creates protein cross links that deactivate digestive enzymes and preserve the cytoskeleton, thereby maintaining cellular morphology.  A side effect of such treatment causes changes in epitopes, making them less accessible to primary antibody binding. High heat treatment in the appropriate pH solution restores epitope configuration allowing for optimal antibody-antigen binding.

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  • Directions for use

  •  Prior to use, dilute buffer to 1X in dd H2O.

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  • Microwave method:

  •  Place slides, held in glass slide rack, in microwavable container, and add enough Epitope Unmasking Solution™ to cover slides completely. Microwave on high until solution comes to a rapid boil, usually ½ to 3 minutes. Turn off microwave. Set microwave to 200W (or  20% power) and simmer for 10 minutes. Remove container from microwave and let sit at room temperature for 20 min. Continue with washing and immunostaining protocol.

  • *** this method should be adjusted for optimal performance for each antibody.

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  • Boiling Method (for fatty tissues, such as brain and breast tissue):

  • Fill a 2L (or larger) glass beaker with 600-750mls 1x Epitope Unmasking Solution™ (enough to cover glass slide rack).  Keep metal handle on. Set on heat plate, add a pinch of boiling chips, and turn heat setting to high. Once boiling, set heat to lowest setting that keeps the solution boiling at a simmer. Add glass slide rack containing samples and boil for exactly 8 minutes. Quickly remove glass beaker and add 500 mls of cold or room temp dd H20. Wait 3 minutes and resume washing step and immunostaining protocol.

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  • Autoclave method ***RECOMMENDED***

  • Fill a 1 liter autoclave resistant plastic container with 300 mls 1X Eptiope Unmasking Solution™. Submerge slides held in glass slide rack, and remove metal handle. Make sure solution totally covers slides. Put in autoclave, set on “slow exhaust” and run at ~121°C for 20 minutes. Remove plastic slide container and keep at room temp in solution for 20 min. Remove slide rack to glass slide holder and continue with washing and Immunostaining protocol.

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  • Storage

  • Store at 4°C. Do not freeze. Discard unused reagent after expiration.

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  • Precautions

  • For professional users.

  • 1.       Follow storage conditions as to minimize the possibility of microbial contamination of reagents.

  • 2.       As with any product, proper handling procedures should   be used.

  • 3.       Wear appropriate protective equipment to avoid contact with eyes and skin.

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  • Notes

  • This product does not contain preservatives. Preservatives such as sodium azide inhibit key enzymes involved in IHC technology such as peroxidases. This quenching can cause the occurrence of false negative results. Due to the absence of preservatives, it is advised to keep strict adherence to storage conditions, proper handling, and disposal of the reagent after expiration occurs.

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  • ***glass slide holder is VWR cat# 25462-006; glass slide rack is VWRcat# 25463-009; Handle is VWR Cat# 25464-001

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