Application
Flow Cytometry
Fixation
Paraformaldehyde
Permeabilization
Yes - 70% methanol
Sample
Human Cell (MCF-7)
Specification
MCF-7
Gating Strategy
live cells
Preparation
Cell harvesting/tissue preparation method: Cell dissociation buffer
Sample buffer: Enzyme free
Sample buffer: Enzyme free
Other product details
Incubation time
1 hour(s) and 0 minute(s) · Temperature: 22°C · Diluent: PBS + 0.2% BSA + 0.1% Sodium azide
Dilution
1/200
Secondary antibody
Name
Non-Abcam antibody was used: goat anti-rabbit IgG PE
Host species: Goat
Clonality: Polyclonal
Conjugation: Phycoerythrin
Host species: Goat
Clonality: Polyclonal
Conjugation: Phycoerythrin
Dilution
1/100
Additional data
Additional Notes
Purple plot represents isotype control, green plot represent Bcl-2 in MCF-7 cells. This antibody did work well in flow, however I am concerned that this might not be recognising Bcl-2, because it detected an ~60kDa protein by western blot.
Abcam response
Thank you for sharing this data with us. ab32124 has previously been tested on Jurkat cells in flow cytometry (please see image on datasheet). However, we are concerned that your western blot data shows a ~60kDa band, which is not specific to Bcl-2. Abcam would like to hear from other customers currently using this product.
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.
Abcam user community
Verified customer
投稿 Sep 02 2014