Alkaline Phosphatase chromogen (BCIP/NBT) - Ready to Use (ab7468)


  • Product name
    Alkaline Phosphatase chromogen (BCIP/NBT) - Ready to Use
    See all Alkaline Phosphatase Chromogen reagents
  • Tested applications
    Suitable for: WB, Dot blot, IHC-P, IHC-Frmore details
  • General notes
    BCIP: 5-bromo-4-chloro-3-indolyl phosphate NBT: p-nitroblue tetrazolium chloride This product used to be called "Alkaline phosphatase detection kit (BCIP/NBT)". Abcam has a wide range of primary and secondary antibodies available on the Abcam website to suit your protocol needs.



Our Abpromise guarantee covers the use of ab7468 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
WB Use at an assay dependent concentration.
BCIP/NBT system is based on hydrolysis of BCIP and reduction of NBT producing a deep purple reaction product. Reaction is several times more sensitive than existing products with little or no background staining. The reaction product is extremely stable and does not fade when exposed to light.
Dot blot Use at an assay dependent concentration.
IHC-P Use at an assay dependent concentration.
IHC-Fr Use at an assay dependent concentration.


This product has been referenced in:
  • Hassani A  et al. Epstein-Barr virus is present in the brain of most cases of multiple sclerosis and may engage more than just B cells. PLoS One 13:e0192109 (2018). Read more (PubMed: 29394264) »
  • Song JW  et al. 3'-UTR engineering to improve soluble expression and fine-tuning of activity of cascade enzymes in Escherichia coli. Sci Rep 6:29406 (2016). WB . Read more (PubMed: 27406241) »

See all 2 Publications for this product

Customer reviews and Q&As

Thank you for your inquiry.

I can confirm that the ab7468 Alkaline Phosphatase chromogen (BCIP/NBT)substrate is ready to use:

Wash the membrane after incubation with the alkaline phosphatase secondary.
Apply sufficient substr...

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Thank you for contacting us.

NBT/BCIP system produces a colorimetric substrate that yields an intense and insoluble black-purple precipitate when reaction with alkaline phosphatase.

Unlike HRP chemiluminiscent methods, the chromogen...

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Thank you for contacting me again. It is always a pleasure to hear from you.

After reviewing your email, I am lead to believe that this really is a hook effect as you had mentioned. I am not an expert on this hook effect in plasma specifical...

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Thank you for contacting us, I am glad to hear that the new product is working well for you.

Usually when color is developing slowly it is one of three issues. 1) the plates and reagents have not come to room temperature. 2) the conjugates a...

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Thank you for contacting us.

Just as with a common indirect ELISA it is important to stop the reaction using the appropriate stop solution.

I hope this information is helpful to you. Please do not hesitate to contact us if you...

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Thank you for contacting Abcam.

Enzymatic detection with an antibody that has been conjugated to HRP or to AP is a widely used and robust method. As this target is located on the peroxisome membrane I would recommend using anAlkaline Phospha...

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Thank you for contacting us.  BCIP/TNBT (ab7413) is similar to BCIP/NBT (ab7468) but is more sensitive and produces a more intense purple color.  Otherwise, the two products will be interchangeable.  I hope this helps, please let me know if you need an...

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Thank you for contacting Abcam regarding ab7468. I contacted the laboratory for the BCIP/NBT concentration, however this information appears to be proprietary. A signal is typically expected within 20 minutes, however we have not specifically teste...

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Thank you for your inquiry. I have contacted the laboratory, however they do not have a protocol for resolubilizing this chromagen. I tried to search the internet, could however not find an adapted protocol for this. I am very sorry for...

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Thank you very much for your enquiry. We do not currently sell substrates such as AP and HRP that have been tested in ELISPOT. This doesn’t mean that these products are not suitable, we just have not tested them in this application. The 4 produc...

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