Go back to the original antibody. If it is no longer supplied then ask abclonal where the original source of the antibody came from and contact that company to buy the one that works well

I recommend reducing the incubation time for the secondary antibody. After incubating with the primary antibody, extend the washing steps with TBST to 10 minutes each. According to the manual, Santa Cruz antibodies should not be frozen, as they are dissolved in gelatin, which can make them inactive or less effective when frozen. I hope this helps.

Hello Gareth Rostro

I had read some reviews of antibodies from Santacruz Biotechnology. I was not happy. They were not so good. Though there may be some good antibodies from Santacruz which I cannot deny.

Attached below are a few links which you may want to refer.

https://www.biocompare.com/Product-Reviews/239545-Poor-quality-ASC-Antibody-antibody-from-Santacruz-Biotech/

https://www.protocol-online.org/biology-forums/posts/17678.html

The quality of antibody matters. Low‐quality antibodies are often the result of poor screening, poor quality control, incomplete characterization and the lack of international standards. For more information you may want to refer to the articles attached below.

https://www.nature.com/articles/521274a

https://www.embopress.org/doi/full/10.15252/embr.202051761#:~:text=Getting%20a%20good%20antibody%20is,the%20lack%20of%20international%20standards.

Article Reproducibility: Standardize antibodies used in research

Article Antibodies: The solution is validation

Chapter Antibodies That Work Again and Again and Again

Article Antibody Validation by Western Blotting

Article Antibody validation: A view from the mountains

I would agree with Paul Rutland. You either go for the original antibody from ABclonal or try another vendor. I would recommend antibodies from Cell Signaling Technology.

Good Luck!