Application
Flow Cytometry
Sample
Rabbit Cell (sHeLa)
Permeabilization
No
Gating Strategy
By size/granularity (FSC-H and SSC-H)
Specification
sHeLa
Preparation
Cell harvesting/tissue preparation method: Collect live cells with cell lifter
Sample buffer: DMEM Life Technologies, 21063-029, used for washes after collection
Sample buffer: DMEM Life Technologies, 21063-029, used for washes after collection
Fixation
None - live cells
Other product details
Incubation time
30 minute(s) · Temperature: 4°C · Diluent: DMEM as above
Dilution
1/100
Secondary antibody
Dilution
1/500
Name
Non-Abcam antibody was used: Goat anti-rabbit IgG (H+L)- 647, A-21244
Host species: Goat
Clonality: Polyclonal
Conjugation: Alexa Fluor 647
Host species: Goat
Clonality: Polyclonal
Conjugation: Alexa Fluor 647
Additional data
Additional Notes
Used to try and detect a DHFR fusion construct localised on the cell surface.
Antibody does not detect construct by flow cytometry. Construct without DHFR was used a control and the signal was the same. Signal in this control was higher than secondary-only control suggesting a high degree of non-specific binding.
Abcam response
Thank you for your valuable feedback about the use of ab49881 in Flow Cytometry. As the antibody had not yet been tested in this application, we were unsure what the result might be and we appreciate the time and effort that the reviewer has put in to testing ab49881 in this application. We encourage all researchers to contact us should they wish to test any of our products in untested species or applications, as these may qualify for our AbTrial program.
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.
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提交于 Oct 31 2017