Application
Flow Cytometry
Sample
Mouse Cell (Dorsal Root Ganglion (DRG) Cells E12)
Permeabilization
Yes - Saponin
Gating Strategy
1. FSC-A/SSC-A (exclusion of cell debris), 2. SSC-W (exclusion of cell doublets)
Specification
Dorsal Root Ganglion (DRG) Cells E12
Preparation
Cell harvesting/tissue preparation method: Trypsinization. Trituration.
Sample buffer: PBS BSA 1%
Sample buffer: PBS BSA 1%
Fixation
Paraformaldehyde
Other product details
Incubation time
16 hour(s) and 0 minute(s) · Temperature: 4°C · Diluent: PBS Saponin 1%
Secondary antibody
Dilution
1/1000
Name
Non-Abcam antibody was used: Alexa Fluor 568 donkey anti-rabbit IgG
Host species: Donkey
Clonality: Polyclonal
Conjugation: Alexa Fluor® 568
Host species: Donkey
Clonality: Polyclonal
Conjugation: Alexa Fluor® 568
Additional data
Additional Notes
GFRa3 expression in DRG cells is not detectable by flow cytometry using the Abcam Ab8028 antibody at dilutions 1:100 and 1:500, as compared to control-incubated DRG cells.
Subsets of E12 DRG neurons express however GFRa3, as judged by immunofluorescence labeling on cryosections of mouse embryos.
Detection of receptors to neurotrophic factors in neurons by using flow cytometry is challenging. The above rating of the ab8028 antibody as "poor" thus relates to its failure in this specific application rather than to its intrinsic quality. Nota bene that the antibody had not been validated for flow cytometry by the producer, therefore this test.
The flow cytometry analyses were done in collaboration with Sébastien Schaller.
Abcam response
Our Scientific Support team is currently investigating this negative Abreview, and we will resolve the issue in accordance with our Abpromise.
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.
Georg Haase
Verified customer
投稿 Oct 14 2015