Thank you for your enquiry.

I am sorry to confirm that ab133406 is not suitable for detection of AP. This detection system has been designed for optimal detection of HRP, and will not be as sensitive for detection of AP.

I am sorry I am not sure which application you are using, but the following two detection substrate kits are suitable for western blotting, and should also be suitable for IHC:

ab7413 Alkaline Phosphatase chromogen (BCIP/TNBT) https://www.abcam.com/Alkaline-Phosphatase-chromogen-BCIPTNBT-ab7413.html https://www.abcam.com/Alkaline-Phosphatase-chromogen-BCIPTNBT-ab7413.html.

ab7468 Alkaline Phosphatase chromogen (BCIP/NBT) https://www.abcam.com/NBT--BCIP-Solution-Ready-to-Use-Alkaline-Phosphatase-chromogen-ab7468.html https://www.abcam.com/NBT--BCIP-Solution-Ready-to-Use-Alkaline-Phosphatase-chromogen-ab7468.html.

I can recommend to review the online datasheets carefully for further information.

I hope this information will be helpful to you. If you have any further questions, please do not hesitate to contact us.

Thank you for contacting us.

As requested, I have attached the Certificates of Compliance for the antibodies and reagents you mentioned - to this email.

I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

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Thank you for confirming the order number.
This is to let you know that I have confirmed and agreed on placing a new order for your customer - for one vial of ab64268 (new and different Batch:GR71729-1) as a free of charge replacement. I hope the second vial will work as it is expected, and please do let me know how you are getting on with this product.
Dear Colleague,
Please add 1 vial of ab64268 (Batch:GR71729-1) to the next shipment for Sapphire Bioscience Pty Ltd.
Original CCE ID: 3445319
Original Order: 948346
Problem is: ALP is inactive and no signal in IHC with this reagent
Many thanks.

Thank you for getting back to me and for re-testing the ALP (AP) activity with new and old chromogen solution. Your co-operation and understanding in this matter is very much appreciated!
I can certainly replace the faulty product (ab64268) as soon as possible.
Once again I apologize for any inconvenience caused and thank you for bringing this matter to our attention.

Thank you very much for getting back to me promptly and for checking the ALP (AP) activity.
This is very important piece of detail and it may indeed imply that AP is not working properly. As I understand Streptavidin Alkaline Phosphatase (ab64268) was purchased from Abcam via the Distributor as well. Would you be so kind to confirm the batch number and the order number of this item as well?
Could you please let Sapphire and myself know how you wish to proceed with this enquiry?
I look forward to hearing from you soon.

Thank you very much for confirming the order details.
Could you please comment on if the ALP (AP) enzyme conjugated to the antibody is inactive? Have you had any chance to check if the ALP is still functioning?
Would you prefer to get a new product or a credit note?
I look forward to hearing from you soon.

Thank you for your enquiry regarding ab7413 and for taking the time to provide some very useful details of the experiments. Your time and effort is much appreciated!
I am very sorry to hear that you are having problems with this chromogen.
As our Abpromise indicates, in the event that a product is not functioning in the applications/species cited on the product data sheet (and the problem has been reported within 6 months of purchase) we will happily offer a credit note/refund to the value of the product purchased.
It could well be that this product has been damaged accidentally either during shipment or storage.
The other alternative would be that the ALP (AP) enzyme conjugated to the antibody is inactive. Have you had any chance to check if the ALP is still functioning?
Would you be so kind to kind the Abcam Order and the date of purchase, our system does not recognize 0761003659 UWA. You may need to contact Sapphire to get the requested information.
I look forward to hearing from you and hope to solve this problem as soon as possible.

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 any additional information. 

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 products identified (ab7413, ab7468, ab64252, ab103742) have been tested in IHC and ab7413 has also been tested in dot blots and WB. This may be a more suitable one to try but unfortunately we cannot guarantee that it would be suitable. I would consider reviewing what ELISPOT kits we have available as we may have one that is suitable. I would be happy to offer any further advice and hope this information helps.

This substrate would detect any endogenous alkaline phosphatase if it were enzymatically active, but I am not sure how activity could remain since alkaline phosphatase is normally inactivated by freezing and chemical treatment during fixation.