MCM3 antibody (ab4460)

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review by Mirjam Eckert submitted on 30 October 2009
Application:	Western blot
MCM3 antibody (ab4460)
Sample	Human Cell lysate - whole cell (HeLa)
Loading amount:	25 µg
Specification	HeLa
Gel Running Conditions:	Reduced Denaturing (12%)
Blocking step:	Milk as blocking agent for  1 hour  · Concentration: 5 % · Temperature: RT°C
Primary Antibody
Dilution	1/2000
Incubation time:	14 hours  Temperature: 4°C Diluent: PBS Tween 5% Milk
Secondary Antibody
Name:	Non-Abcam Antibody was used: Jackson ImmunoResearch Conjugation: HRP
Dilution:	1/10000
Detection
Detection method:	ECL
Exposure:	10 seconds
Bands:	Specific: 105 kDa   Non-specific: 50, 65 kDa
Additional Data
Rating	Good
Additional notes	RIPA lysis buffer, nitrocellulose membrane, molecular weight markers 100, 75, 50kDa.
Image	MCM3 antibody for WB (Human) Copyright image used under license

review by Madlaina Breuleux submitted on 28 July 2004
Application:	Western blot
MCM3 antibody (ab4460)
Additional Data
Notes	I use the antibody against mcm3 to have a loading control for western blot analysis; this works very nicely for nuclear cell extracts as well as for SDS cell extracts!
Dilution notes	1:2000 in 5% milk/PBS The milk can be stored at -20?C and re-used for about 5-8 times! (the antibody is very specific)

review by Anne Early submitted on 16 April 2004
Application:	Western blot
MCM3 antibody (ab4460)
Additional Data
Notes	Works well in Western, recognizes a single band, no background problems. Have used as a 1:2000 dilution.
Dilution notes	1:2000 dilution

review by Viviane Bechtold submitted on 11 December 2003
Application:	Immunoprecipitation
MCM3 antibody (ab4460)
Additional Data
Notes	- Applications used: We used this antibody for Western Blot analysis and Immunoprecipitation of MCM3 proteins from nuclear extracts - Recommended dilutions: WB: 1/2000 (optimal dilution in our hands) IP: 1ug of the antibody per 50ug Lysat - Tissue or cell-line used: Keratinocyte cell lines like W12 cells or S12 cells (= equivalent to HeLa cells) - Detection system used: ECL detection kit (Amersham) - How well it worked: We used the dilutions described earlier corresponding to the optimal values for each experiment in our hands. It worked very well and we needed maximum 1mn exposition to detect the protein on the film. WB: W12 or S12 nuclear extract (50ug) incubated with abcam MCM3 antibody used at 1/2000for 1 hour IP: S12 nuclear extract (50ug) incubated with ABCAM MCM3 antibody using at 1ug/50ug lysat and overnight incubation. - Any special conditions: We did the IP from nuclear extracts and not from a total cell extraction
Dilution notes	WB: 1/2000 IP: 1ug per 50ug of nuvlear extract