BCIP-NBT Dye for IHC

Cat. No.
G447
Unit
250 ml
Price
$205.00
Cat. No. G447
Name BCIP-NBT Dye for IHC
Unit 250 ml
Category Gels, Blots & ELISAs
Description

When 5-Bromo-4-Chloro-3-Indolyl Phosphate (BCIP)/Nitroblue Tetrazolium (NBT) is added to tissue sections containing Alkaline Phosphatase (AP) labeled probes, the hydrolysis of the BCIP phosphate group by AP results in the formation of powerful reduced substances. These substances reduce NBT to insoluble NBT formazan products, which is readily visible as fine purple deposits at sites of AP activity. ABM’s BCIP/NBT Dye for Immunohistochemistry provides the greatest sensitivity amongst the immunohistochemistry dyes.

Application

Suitable for the immunohistochemical localization of Alkaline Phosphatase (AP) labeled probes. This dye may also be used for In Situ hybridization and blotting techniques.

Recommended Protocol 1. Complete all required incubations with antibodies, probes, and AP labeled reagents. 2. Wash tissues sample with Tris-HCl or Tris buffered saline containing 0.1% Tween 20. Note: DO NOT USE phosphate buffers. Inorganic phosphate is a powerful inhibitor of Alkaline Phosphatase. 3. After the final wash, completely cover the sample in the dye solution and incubate for 5-15 minutes. Protect the reaction from light. 4. Stop the enzyme reaction by thoroughly washing the sample with dH2O. Presence of excessive background staining indicates incomplete removal of non bound AP. Increase the number of washes or the washing time to remedy the problem. 5. If desired, counterstain the sample using a 0.5% solution of neutral red prepared in a 0.1M acetate buffer, pH 5.2.
Storage Condition

Store at room temperature (17-28ºC). Storage at refrigerator temperatures (2-4ºC) will not inhibit product performance but warm the product to reaction temperatures prior to use. Discard if solution turns purple or turbid.

Material Citation If use of this material results in a scientific publication, please cite the material in the following manner: Applied Biological Materials Inc, Cat. No. G447
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