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BCIP/NBT 203790

点击次数：195发布时间：2016/9/30 15:16:43

更新日期：2023/10/7 10:42:25

所在地：中国大陆

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简单介绍：BCIP/NBT 203790 ，Immunoblotting (see comments)

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Description
Overview	Ready-to-use alkaline phosphatase substrate solution. Suitable only for immunoblotting.
Catalogue Number	203790
Brand Family	Calbiochem®
Synonyms	5-Bromo-4-chloro-3-indolylphosphate/Nitroblue Tetrazolium

References
References	De Jong, A.S., et al. 1985. Histochem. J. 17, 1119.

Product Information
Form	Clear to pale yellow liquid
Formulation	0.577 mM BCIP, 0.122 mM NBT in a proprietary solution.
Preservative	None

Applications
Application Notes	Immunoblotting (see comments)
Application Comments	It is not recommended for immunohistochemistry or in situ hybridization. Functionality check performed on final product by dot blot. Additional QC testing is preformed on a lot-specific basis: pH: 9.7-9.9 Stability at 4°C Stability at 18-26°C Suggested procedure for immunoblot staining with BCIP/NBT: 1. Complete all required incubations with antibodies ａnd alkaline-phosphatase-labeled probes. 2. After the final binding reaction with the alkaline-phosphatase labeled probe, wash the membrane thoroughly in a buffer such as Tris-buffered saline (TBS) containing 0.1% Tween®-20 detergent. DO NOT USE PHOSPHATE BUFFERS; inorganic phosphate is a potent inhibitor of alkaline phosphatase. 3. Following the final wash, completely cover the membrane with BCIP/NBT Solution ａnd incubate, protected from light, for 5-15 min, or until the desired color intensity is obtained (dark purple bands or dots will appear at the sites of enzyme activity). Note: Variables associated with assay conditions will dictate the proper reaction time. If the color develops almost immediately, the NBT-formazan deposit may flake off the membrane. If this occurs, further dilution of the alkaline phosphatase probe is recommended. A fine line of formazan deposit circumscribing the band or dot, with no deposit in the center, also suggests that further dilution of the alkaline phosphatase probe is needed. 4. Stop the reaction by washing the membrane thoroughly in distilled water. Note: There should be little or no background. Presence of excessive background staining indicates incomplete removal of unbound alkaline phosphatase from the membrane. To remedy this problem, increase the number of wash steps or washing times. 5. Air-dry membranes ａnd store at room temperature, protected from light.

Applications

Application Notes

Immunoblotting (see comments)

Application Comments

It is not recommended for immunohistochemistry or in situ hybridization. Functionality check performed on final product by dot blot. Additional QC testing is preformed on a lot-specific basis:
pH: 9.7-9.9
Stability at 4°C
Stability at 18-26°C

Suggested procedure for immunoblot staining with BCIP/NBT:

1. Complete all required incubations with antibodies ａnd alkaline-phosphatase-labeled probes.
2. After the final binding reaction with the alkaline-phosphatase labeled probe, wash the membrane thoroughly in a buffer such as Tris-buffered saline (TBS) containing 0.1% Tween®-20 detergent. DO NOT USE PHOSPHATE BUFFERS; inorganic phosphate is a potent inhibitor of alkaline phosphatase.
3. Following the final wash, completely cover the membrane with BCIP/NBT Solution ａnd incubate, protected from light, for 5-15 min, or until the desired color intensity is obtained (dark purple bands or dots will appear at the sites of enzyme activity).
Note: Variables associated with assay conditions will dictate the proper reaction time. If the color develops almost immediately, the NBT-formazan deposit may flake off the membrane. If this occurs, further dilution of the alkaline phosphatase probe is recommended. A fine line of formazan deposit circumscribing the band or dot, with no deposit in the center, also suggests that further dilution of the alkaline phosphatase probe is needed.
4. Stop the reaction by washing the membrane thoroughly in distilled water.
Note: There should be little or no background. Presence of excessive background staining indicates incomplete removal of unbound alkaline phosphatase from the membrane. To remedy this problem, increase the number of wash steps or washing times.
5. Air-dry membranes ａnd store at room temperature, protected from light.

Storage ａnd Shipping Information
Ship Code	Ambient Temperature Only
Toxicity	Standard Handling
Storage	+2°C to +8°C
Protect from Light	Protect from light
Do not freeze	Ok to freeze
Special Instructions	Discard if the solution turns purple or turbid. The reagent is stable at room temperature, but we recommend storage in the refrigerator to increase the shelf life. Warm to assay temperature before use.

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点击次数：195发布时间：2016/9/30 15:16:43

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