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ReviewID: xxx AbID: 25273, Anti-Ly6g antibody [RB6-8C5] (Allophycocyanin) Rating: Poor Image: Anti-Ly6g antibody [RB6-8C5] (Allophycocyanin) for Flow Cytometry (Dog) Sample: Species: Dog Type: Cell Specification: from whole blood Preparation: Cell harvesting/tissue preparation method: after red blood cell lysis
Sample buffer: PBS/BSA 1% Application: Application: Fixation: Paraformaldehyde Permeabilization: No Gating Strategy: alive cells Other detail: Dilution: 1/100 Incubation time: 1 hour at 4°C Diluent: PBS/BSA 1% Secondary Antibody: Name: None used Conjugation: Additional Data:
Asked on Nov 07 2012
Thank you for submitting an Abreview for ab25273. 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 (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:
- fix with PFA after incubation with the primary antibody - if you haven't done that already.
- incubate longer with the primary antibody (e.g. 2-3 h at RT or overnight at 4 degree)
It could also just be the case that the antibody does not work for dog samples due to low homology.
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 Nov 07 2012