Anti-MMP14 antibody (ab3644)

  Western blot - MMP14 antibody (ab3644)

All lanes : Anti-MMP14 antibody (ab3644) at 1/500 dilution

Lane 1 : 15ug MCF-7 whole cell lysate
Lane 2 : 15ug MSTO-2H11 whole cell lysate
Lane 3 : 15ug Malme-3M whole cell lysate
Lane 4 : 15ug Calu-1 whole cell lysate
Lane 5 : 15ug Sk-Lu-1 whole cell lysate

Secondary
Peroxidase-conjugated affinipure goat anti-rabbit
developed using the ECL technique

Performed under reducing conditions.

Observed band size : 54,66 kDa (why is the actual band size different from the predicted?)
Additional bands at : 20 kDa (possible non-specific binding),98 kDa (possible non-specific binding).

Exposure time : 30 seconds

This image is courtesy of an anonymous Abreview

See Abreview

  Immunocytochemistry/ Immunofluorescence-MMP14 antibody(ab3644)

ICC/IF image of ab3644 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 (ab3644, 1µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit 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.

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MMP14 antibody (ab3644)

IHC image of ab3644 staining in Hu Colon formalin fixed paraffin embedded tissue section, performed on a Leica Bond^TM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab3644, 5µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.