Anti-MCM7 antibody [47DC141] (ab2360)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - MCM7 antibody [47DC141] (ab2360)

ab2360 - immunohistochemistry

Formalin fixed paraffin embedded human tonsil stained with MCM7, using ABC and DAB chromagen.

Immunocytochemistry/ Immunofluorescence - MCM7 antibody [47DC141] (ab2360)

This image shows immunostaining of rat brain endothelial cells. Brain endothelial cells were co-cultured with neuronal precursor cells and the nuclear staining represents cells in cell cycle. Primary antibody (ab2360) was used at 1:50 dilution, incubated overnight at 4 ^oC.  Secondary antibody - Alexafluor (488 nm) at 1:200 dilution, incubated for 2 hours at room temperature.

The picture was kindly supplied by Dr Joseph Corteza Lim and Dr Margery Barrand from University of Cambridge, Department of Pharmacology.

Western blot - MCM7 antibody [47DC141] (ab2360)

All lanes : Anti-MCM7 antibody [47DC141] (ab2360) at 1/200 dilution

Lane 1 : M phase Xenopus laevis egg extract, whole tissue lysate.
Lane 2 : I phase Xenopus laevis egg extract, whole tissue lysate.

Secondary
HRP conjugated Donkey anti-rabbit IgG
developed using the ECL technique

Predicted band size : 80 kDa
Observed band size : 95 kDa (why is the actual band size different from the predicted?)


Exposure time : 1 minute

This image is courtesy of an anonymous Abreview

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - MCM7 antibody [47DC141] (ab2360)

ab2360 staining MCM7 in human bladder cancer tissue sections by Immunohistochemistry (formalin fixed sections). Tissue was fixed with formaldehyde and a heat mediated antigen retrieval step was performed using citrate buffer. Tissue was blocked with 5% BSA for 1 hour at room temperature followed by incubation with the primary antibody at a 1/1200 dilution for 1 hour. A HRP-conjugated goat anti-mouse polyclonal was used as secondary antibody un-diluted.

Image kindly supplied by Dr Karin Birkenkamp-Demtroeder through Abreview

Western blot - MCM7 antibody [47DC141] (ab2360)

Lane 1 : Anti-MCM7 antibody [47DC141] (ab2360) at 1/50 dilution
Lane 2 : Anti-MCM7 antibody [47DC141] (ab2360) at 1/200 dilution
Lane 3 : Anti-MCM7 antibody [47DC141] (ab2360) at 1/500 dilution

Lane 1 : Whole cell lysate prepared from SW780 cells
Lane 2 : Whole cell lysate prepared from SW780 cells
Lane 3 : Whole cell lysate prepared from SW780 cells

Lysates/proteins at 25 µg per lane.

Secondary
Goat anti-mouse IgG conjugated to HRP at 1/5000 dilution
developed using the ECL technique

Performed under reducing conditions.

Predicted band size : 80 kDa
Observed band size : 81 kDa (why is the actual band size different from the predicted?)


Exposure time : 10 minutes

Gel run under denaturing conditions 4-12% gradient.

Image kindly supplied by Dr Karin Birkenkamp-Demtroeder through Abreview

Immunocytochemistry/ Immunofluorescence-MCM7 antibody [47DC141](ab2360)

ICC/IF image of ab2360 stained HeLa cells. The cells were 4% formaldehyde fixed (10 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 (ab2360, 1µg/ml) overnight at +4ºC. The secondary antibody (green) was Alexa Fluor® 488 goat anti-mouse IgG (H+L) 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.

Flow Cytometry-MCM7 antibody [47DC141](ab2360)

Overlay histogram showing HeLA cells stained with ab2360 (red line). The cells were fixed with 100% methanol (5 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 (ab2360, 1/20 dilution) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. 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.