Question (44255) | Anti-CACNA1C antibody [L57/46] (ab84814)

Go to datasheet (ab84814)

Question

ReviewID: xxx AbID: 84814, Anti-DHPR alpha 1 antibody [S57-46] Rating: Inconclusive Image: Anti-DHPR alpha 1 antibody [S57-46] for Immunohistochemistry (Frozen sections) (Human) Sample: Species: Human Type: Tissue sections Specification: atrium Application: Application: Fixative: Paraformaldehyde Permeabilization: Yes Blocking step: BSA as blocking agent for 1 hour at 25°C Other detail: Dilution: 1/50 Incubation time: 12 hours at 4°C Diluent: PBS+BSA+serum Secondary Antibody: Name: Non-Abcam Antibody was used: goat anti mouse IgG2b488 Conjugation: Alexa Fluor® 488 Dilution: 1/500 Additional Data: Additional Notes: only see non-specifc binding

Answer

Thank you for submitting an Abreview of ab84814. As you may have noticed, your review has now been published on our website.

Since you obtained poor results using the antibody in a tested species and application, 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) Trying another fixative might help to solve the problem as the PFA fixation might crosslink antibody binding sites.
2) You could also try omitting thepermeabilization step if your sections are thin enough (10 um or less).
3) Doing ano primary control could help to determine if the secondary antibody contributes to the non-specific staining.
4) What is you order or PO number?

If you decide to follow these suggestions, please let me know if they are helpful. Additionally, this antibody is covered by our Abpromise, so if the suggestions do not help and the antibody was purchased within the past 6 months, I would be happy to discuss a refund or replacement with you to resolve this issue.

I look forward to your reply.

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