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ReviewID: xxx AbID: 133211, Anti-alpha 1 Sodium Potassium ATPase antibody Rating: Inconclusive Image: Anti-alpha 1 Sodium Potassium ATPase antibody for Immunocytochemistry/ Immunofluorescence (Mouse) Sample: Species: Mouse Type: Cell Specification: Liver Application: Application: Fixative: Formaldehyde Permeabilization: Yes Blocking step: Serum as blocking agent for 1 hour at 25°C Other detail: Dilution: 1/20 Incubation time: 16 hours at 4°C Diluent: 1% donkey serum Secondary Antibody: Name: Non-Abcam Antibody was used: Alexa Fluor 488 Conjugation: Alexa Fluor® 488 Dilution: 1/1000 Additional Data:
Asked on Oct 18 2012
Thank you for submitting an Abreview for ab133211. As you may have noticed, your review has now been published on our website.
Since you obtained poor results using the antibody in an untested species, 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:
- The image you provided does show membrane staining of the antibody.
- I might suggest using less primary antibody (e.g. 1/50) as well as reducing the incubation time to several hours at room temp - in order to make the signal more specific and reduce some background (if you have not tried that already).
If you decide to follow these suggestions, please let me know if they are helpful.
I wish you good luck with your research.
Answered on Oct 18 2012