Flow Cytometry abreview for Anti-Bcl-2 antibody [E17]

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

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
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.

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Submitted Sep 02 2014