Apc5 antibody (ab4170)

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review by Kirk McManus submitted on 31 October 2005
Application:	Immunocytochemistry
Apc5 antibody (ab4170)
Sample	Human Cell (HeLa, SK-N-SH and Indian muntjac)
Specification	HeLa, SK-N-SH and Indian muntjac
Fixative:	Paraformaldehyde
Primary Antibody
Dilution	1/100
Incubation time:	30 minutes
Secondary Antibody
Name:	Non-Abcam Antibody was used: Goat Anti-Rabbit IgG (H+L) Conjugation: Cy3 ®
Dilution:	1/200
Additional Data
Rating	Good
Additional notes	I tested this antibody in paraformaldehyde fixed HeLa and SK-N-SH cells and it appeared to work best at 1:100 dilution in both cell lines. This antibody gives a focal staining pattern during the middle stages of mitosis (i.e. prometaphase to anaphase), where the foci localize to regions around the chromosomes. Cells were fixed in 4% paraformaldehyde for 10min. Cells were permeablized in 0.5% Triton-X100 in PBS for 10min and then rinsed 3x in PBS. Cells were incubated with a 1:100 dilution of antibody in PBS for 30min, rinsed 1x in 0.1% Triton-X100 and rinsed 3x in PBS prior to a 30min incubation with an appropriate secondary antibody (1:200). Cells were then rinsed 1x in 0.1% Triton-X100 and 2x in PBS before being mounted using a 90% glycerol/PBS mounting media containing both DAPI and PPDA (anti-fade reagent). I also tested this antibody for reactivity in Indian muntjac (IM) cells and the staining pattern was similar to that of HeLa and SK-N-SH. The accompanying image is of a metaphase IM cell. The DNA is stained with DAPI (blue) and the APC5 is stained green.
Image	Apc5 antibody for ICC (Human) Copyright image used under license
Abcam response	Thank you for your review of ab4170 and the accompanying images. We are happy to hear that this antibody has worked successfully in this application and as a result we are able to upgrade this antibody from Fast Track status.