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Synthetic peptide corresponding to Human Cyclin E1.
This antibody may be diluted to a titer of 1:100 - 1:200 with an enzyme-polymer detection method. The Final dilution sould be determined by the user based upon the staining conditions employed.
Our Abpromise guarantee covers the use of ab3927 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|Flow Cyt||1/10 - 1/50.
ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.
|ICC/IF||1/50 - 1/200.|
|IHC-P||1/100 - 1/200. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
This antibody may be diluted to a titer of 1:100 - 1:200 with an enzyme-polymer detection method.
|IHC-Fr||1/100 - 1/200.|
|WB||1/500 - 1/1000. Detects a band of approximately 42 kDa (predicted molecular weight: 47 kDa).
Detects a doublet of 50 kDa and a single band of 42 kDa, see Abreviews.
Overlay histogram showing MCF-7 cells stained with ab3927 (red line). The cells were fixed with 4% paraformaldehyde (10 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab3927, 1/10 dilution) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1:500 dilution for 30 min at 22°C. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in MCF-7 cells fixed with methanol (5 min)/permeabilized in 0.1% PBS-Tween used under the same conditions.
ICC/IF image of ab3927 stained HepG2 cells. The cells were 100% methanol fixed (5 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab3927, 1/100 dilution) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-mouse IgG (H+L) ab150113 used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.
Immunoblot analysis of Cyclin E expression in NIH/3T3 cells. Cells were transfected with the indicated plasmids and harvested 48 hr post transfection. Cyclin E expression was analysed by immunoblotting with ab3927. Immunoblot analysis of Cyclin E expression in NIH3T3 cells. Cells were transfected with the indicated plasmids and harvested 48 hr post transfection. Cyclin E expression was analysised by immunoblotting with ab3927.
ab3927 staining human breast carcinoma by IHC-P.
ab3927 - immunohistochemistry
Formalin fixed paraffin embedded human breast carcinoma stained with Cyclin E, using ABC and DAB chromagen.