Anti-MMP12 antibody - Carboxyterminal end (ab66157)

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)-MMP12 antibody - Carboxyterminal end(ab66157)

Ab66157 staining human breast carcinoma. Staining is localized to the nuclear membrane and cytoplasm.
Left panel: with primary antibody at 2 ug/ml. Right panel: isotype control.
Sections were stained using an automated system DAKO Autostainer Plus , at room temperature. Sections were rehydrated and antigen retrieved with the Dako 3-in-1 AR buffers EDTA pH 9.0 in a DAKO PT Link. Slides were peroxidase blocked in 3% H2O2 in methanol for 10 minutes. They were then blocked with Dako Protein block for 10 minutes (containing casein 0.25% in PBS) then incubated with primary antibody for 20 minutes and detected with Dako Envision Flex amplification kit for 30 minutes. Colorimetric detection was completed with Diaminobenzidine for 5 minutes. Slides were counterstained with Haematoxylin and coverslipped under DePeX. Please note that for manual staining we recommend to optimize the primary antibody concentration and incubation time (overnight incubation), and amplification may be required.

  Western blot - MMP12 antibody - Carboxyterminal end (ab66157)

All lanes : Anti-MMP12 antibody - Carboxyterminal end (ab66157) at 1 µg/ml

Lane 1 : A549 (Human lung adenocarcinoma epithelial cell line) Whole Cell Lysate
Lane 2 : WI38 (Human lung fibroblast cell line) Whole Cell Lysate

Lysates/proteins at 10 µg per lane.

Secondary
Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution
developed using the ECL technique

Performed under reducing conditions.

Predicted band size : 54 kDa
Observed band size : 54 kDa
Additional bands at : 22 kDa. We are unsure as to the identity of these extra bands.

Exposure time : 12 minutes

  Immunocytochemistry/ Immunofluorescence-MMP12 antibody - Carboxyterminal end(ab66157)

ICC/IF image of ab66157 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 (ab66157, 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.