Streptavidin HRP (ready-to-use) (ab64269)

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ab1002 Anti-Rabies Virus antibody looks very promising as it has been established for the use in IHC-P and should detect a wide range of rabies strains. As a starting dilution the customer can try a 5 ug/ml dilution incubated overnight at 4C. However, it may be that the actual optimum dilution may be lower or higher, but this depends on her samples. Another point is that I would strongly recommend a heat induced Antigen Retrieval if her slides are fixed with formalin.


https://www.abcam.com/index.html?datasheet=1002 (or use the following: https://www.abcam.com/index.html?datasheet=1002).


A matching secondary antibody would be Goat polyclonal Secondary Antibody to Mouse IgG2a - heavy chain (Biotin), pre-adsorbed (ab98696)


https://www.abcam.com/index.html?datasheet=98696 (or use the following: https://www.abcam.com/index.html?datasheet=98696).


To be fair, I am not the biggest fan of ready- to- use solution (andmost of my colleagues here a neither), for the following reason: most of all antibodies need optimization, may it be the blocking step, the retrieval time or method, the antibody incubation buffer etc. With ready-mix solutions it almost impossible to adjust the experiment settings. Ready mix solution make sense if the antibody is used in a high -throughput lab (like 20 slides at once with a fixed protocol), but not if you have to "learn" IHC-P and have the freedom to adjust the protocol fore the perfect results. In addition, it's cheaper ;)


Of course, once the protocol is optimized, using ready mixed solution are a very time efficient and make life easier, but I wouldn't use them as a beginner.


All the buffers your customer needs (see attachment) are listed in our very detailed and easy to follow step-by-step our protocol. However, none of my advice is written in stone, so if your customer prefers the ready mix solutions, we are more than happy to sell them to her.


Something we cannot provide are the reagents for the de-paraffinization, I am afraid, but a company like Sigma should be able to help with that.


Pease sent the customer our protocol and my advice. I am more than happy to assist her to get her experiments running. And another thing: she shouldn't be shy to ask any question she may have about the protocol!

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

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Here is the list with the requested items. However, please be aware that this is not a technical inquiry as all item could be found by using the search application on our website under the "reagents" category. Please let us know if you need help on navigating our website so you can help your customers faster than relying on us.


Regarding the Rabies antibody, I am afraid we do not have what your customer requests: All we have are mouse monoclonal antibodies against this virus, but none produced in goat. Please let me know if this expectable for her, so I can suggest the right primary antibody with the matching biotinylated secondary antibody.


https://www.abcam.com/index.html?datasheet=64214 (or use the following: https://www.abcam.com/index.html?datasheet=64214).

https://www.abcam.com/index.html?datasheet=64216 (or use the following: https://www.abcam.com/index.html?datasheet=64216).

https://www.abcam.com/index.html?datasheet=64238 (or use the following: https://www.abcam.com/index.html?datasheet=64238).

https://www.abcam.com/index.html?datasheet=64269 (or use the following: https://www.abcam.com/index.html?datasheet=64269).

https://www.abcam.com/index.html?datasheet=94666 (or use the following: https://www.abcam.com/index.html?datasheet=94666).

https://www.abcam.com/index.html?datasheet=128988 (or use the following: https://www.abcam.com/index.html?datasheet=128988).

https://www.abcam.com/index.html?datasheet=128990 (or use the following: https://www.abcam.com/index.html?datasheet=128990).



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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We do not currently offer double staining IHC kits. Your staining conditions are challenging as, of the two common HRP chromogens, only DAB is alcohol insoluable. While of the common AP chromagens, only BCIP/NBT is insoluable. This means that to do double labeling you will have to do a sequential stain first using one detection system and then with the other.

I have collected a list, with links, of the reagents which you would need for this type of staining.

1) After de-parrafinizing and re-hydrating the samples, perform an antigen retrieval step treating with Cirate buffer pH6.0,https://www.abcam.com/10x-Citrate-Buffer-pH-6-0-ab64214.html, in the microwave or pressure cooker. After performing the antigen retrieval rinse in cold tap water for 10 minutes.

2) Rinse slides in TBST,https://www.abcam.com/20x-TBS-T-with-Tween-20-ab64250.html.

3) Performing a blocking step using protein block,https://www.abcam.com/Protein-Block-ab64226.html.

4) Incubate sections with your first primary antibody, for clarity I will assume the CD44 antibody, raised in mouse.

5) Wash with TBST.

6) Block endogenous peroxidases using aHydrogen Peroxide Blocking Reagent,https://www.abcam.com/Hydrogen-Peroxide-Blocking-Reagent-ab94666.html.

7) Incubate your tissue with a biotinylated secondary antibody, in this instance anti-Mouse IgG,https://www.abcam.com/Biotinylated-Goat-anti-Mouse-IgG-H-L-Ready-to-Use-ab64255.html.

8) Wash with TBST.

9) Detect your first antibody usingStreptavidin Peroxidase (Ready to Use)https://www.abcam.com/Streptavidin-Peroxidase-Ready-to-Use-ab64269.html. Note that this does not containa serum or BSA solution as diluent. Serum may contain biotin therefore competing Streptavidin binding with biotinylated secondary antibody, therefore reducing binding activity.

10) Apply your chromogen, DABsubtrate,https://www.abcam.com/DAB-Substrate-Kit-ab64238.html.

11) Wash with TBST.

12)Performing a second blocking step using protein block,https://www.abcam.com/Protein-Block-ab64226.html.

13)Wash with TBST.

14)Incubate sections with the your second primary antibody, this time ALDH1, raised in rabbit.

15)Wash with TBST.

16)Incubate your tissue with a biotinylated secondary antibody, in this instance anti-Rabbit IgG,https://www.abcam.com/Biotinylated-Goat-Anti-Rabbit-IgG-H-L-Ready-to-use-ab64256.html

17)Wash with TBST.

18)Detect your first antibody usingStreptavidin Alkaline Phosphatase (Ready to Use)https://www.abcam.com/Streptavidin-Alkaline-Phosphatase-Ready-to-Use-ab64268.html.

19) Apply your second chromogenAlkaline Phosphatase chromogen (BCIP/NBT) - Ready to Use,https://www.abcam.com/Alkaline-Phosphatase-chromogen-BCIP-NBT-Ready-to-Use-ab7468.html.

18) Wash with TBST, counterstain if desired.


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

Merci de nous avoir contactés.

Les deux kits que je vous recommende sont :

Streptavidin Alkaline Phosphatase (Ready to Use), www.abcam.com/ab64268
Streptavidin Peroxidase (Ready to Use), www.abcam.com/ab64269



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