Anti-Substance P antibody [SP-DE4-21] (ab14184)

Thank you for contacting us.

We haven't specifically tested human brain with this antibody, but I think it can be used as a positive control. According to the Human Protein Atlas, there is low moderate staining in human brain.

http://www.proteinatlas.org/ENSG00000006128/normal

Plus, one of our other anti-substance P antibodies has been tested in human brain.

https://www.abcam.com/Substance-P-antibody-EPR3959-ab133240.html

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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I am very pleased to hear your customer would like to test ab14184 Anti-Substance P antibody [SP-DE4-21] in rabbit.This code will give them 1 free primary antibody before the expiration date. To redeem this offer, please submit an Abreview for rabbit and include this code in the “Additional Comments” section so we know the Abreview is for this promotion. For more information on how to submit an Abreview, please visit the site: www.abcam.com/Abreviews.


Remember, we publish both positive and negative Abreviews on our datasheets so please submit the results of your tests. The code will be active once the Abreview has been submitted and can be redeemed in one of the following ways: 1) Call to place your order and mention the code to our customer service department; 2) Include the code in your fax order; 3) Place your order on the web and enter the promotional code.

Any feedback that you can provide will be greatly appreciated, whether positive or negative. If you have any further questions, please do not hesitate to contact us. We look forward to receiving your Abreview and wish you luck with your research.

The terms and conditions applicable to this offer can be found here: www.abcam.com/collaborationdiscount.

Thank you for your reply.

As I had mentioned, we have not validated ab14184 for use on frozen sections. Not all antibodies will work for every application, and for that reason we only cover tested applications and species under our Abpromise guarantee.

While antibodies that have been validated for use on paraffin-embedded sections may work with frozen sections, they are not always compatible. The main difference between these assays is the use of antigen retrieval. For any formaldehyde-based fixation performed for longer than 20 minutes, the proteins will be cross-linked. If the epitope recognized by the antibody is obscured by the methylene bridges formed by formaldehyde fixation, no specific binding will be observed. Antigen retrieval will partially digest the methylene bridges and expose more epitopes in the protein. While this is important for polyclonal antibodies, it can be especially crucial for monoclonal antibodies that recognize only one epitope. Our Substance P antibody ab14184 has not been tested on cross-linked samples without the use of an antigen retrieval step.

If you are ever interested in testing an antibody in an untested species or application, please feel free to inquire about our 100% Testing Discount prior to purchase:

https://www.abcam.com/collaborationdiscount

I would be happy to help you troubleshoot this further to improve your IHC-Fr results, or to offer you a refund or credit if the antibody does not work in one of our tested applications: ELISA, IHC-P, or ICC/IF. Please let me know if you have any further questions or concerns.

Thank you for contacting us. I am sorry that ab14184 is giving non-specific staining in IHC-Fr. We have not validated this antibody for use on frozen sections, but it should be suitable for that purpose with some troubleshooting.

1) What species did your samples come from?

2) Could you please let me know the dilutions and incubation times used for the primary and secondary antibodies? Have you performed a no-primary antibody control to rule out the secondary antibody as the cause of the background staining? It may help to reduce the secondary antibody concentration to around 1:500 - 1:1000.

3) Were the sections fixed with 4% PFA for 20 minutes or just cryoprotected with the sucrose treatment? We have been able to obtain staining using a 4% PFA fixation with Proteinase K antigen retrieval or heat mediated antigen retrieval. It may help to add a 10 minuteProteinase K treatment to your protocol prior to blocking. Alternatively, you could perform the sucrose cryoprotection followed by methanol fixation.

4) To ensure that the excess antibody is washed off, I would recommend doing 3 x 5 minute washes in PBST.

5) Since you appear to be getting a lot of background staining, you could try blocking with 10% normal goat serum for a longer period of time. Usually30 minutes to an hour is sufficient, butincubating overnight with the blocking agent canhelp to further reduce the background.

I hope this helps, if not, please let me know and I will be happy to help you further.

Thank you for contacting us.


I performed a protein alignment with the target immunogens and the rabbit protein sequences for the antibodies. We recommend that alignment should be over 85% to predict that an antibody will detect in a different species.

Here are the results, similarities are in %:

ab47027 Anti-CGRP antibody: 85%

ab14184 Anti-Substance P antibody [SP-DE4-21]: 100%

ab106291 Anti-Substance P antibody: 100%

ab2318: As mentioned in the Scientific Support section, PGE2 is a lipid rather than a protein and is therefore not species specific. There should not be any particular problems using this assay to detect PGE2 from rabbit. Optimization for different sample types will have to be done by the end user. Please keep in mind that since PGE2 is a lipid, extraction buffers containing detergents (for preparing tissue samples) are not recommended. Mechanical disruption in a buffer such as PBS is recommended instead. For serum and plasma, simple dilution should be sufficient.


I can offer a discount off a future purchase if your customer buys one or more of the antibodiesnow, test it in rabbit and submit feedback to us in the form of an Abreview. It doesn’t matter whether the Abreview is positive or negative, we would just really like to receive your feedback.


If they are interested in this offer, please let me know.

Thank you for contacting us and reporting the formatting errors on the datasheet of Anti-Substance P antibody (ab14184).

I can confirm that the antibody was used to stain PC12 cells with a concentration of 1 µg/ml. This information has now been updated on the datasheet of ab14184.

I hope this information has been of help. If you require any further information please do not hesitate to contact us again.

Thank you for your reply.

This is nevertheless good information to have. I made a note of it in our system.

Thank you so much for trying the antibody in dog samples!

Please do not hesitate to contact us if you need any more advice or information.

Thank you for submitting an Abreview of ab14184. As you may have noticed, your review has now been published on our website.

Since you obtained poor results using the antibody in a untested species (dog), we would like to follow up on this to see if we can possibly improve the results you are seeing with this antibody.

Based on the information in your review, I feel that altering the following step in your protocol may help improve your results:

1) Try another antigen retrieval step (e.g. citrate buffer or Tris/EDTA).
2) Try using more primary antibody.

If you decide to follow these suggestions, please let me know if they are helpful.

I wish you good luck and look forward to your reply.