Western blot - MMP9 antibody (ab38898)

Anti-MMP9 antibody - Whole molecule (ab38898) + Human MMP9

Observed band size : 88,92 kDa (why is the actual band size different from the predicted?)


Ab38898 detects a band at 92 Kd (pro-form) and a band at 88 Kd (active form). Mouse MMP9 is slightly larger than human MMP9, and the antibody detects a band at about 105 Kd. It is recommend to concentrate samples by Gelatin-agarose affinity chromatography prior to Western blot usage. A recommended starting concentration for Western blots is 1:1000 when using colorimetric substrates such as BCIP/NBT, and 1:5000 for chemiluminescent substrates. Higher concentration of antibody may be needed for non-human samples.

Immunohistochemistry (Frozen sections) - MMP9 antibody - Whole molecule (ab38898)

ab38898 at a 1/1000 dilution staining mouse heart tissue by Immunohistochemistry (Frozen sections). The tissue was removed from a mouse, rinsed in PBS and slowly frozen in supercooled isopentane. 14um sections were made using a cryostat. The sections were acetone fixed and blocked in 2% BSA prior to incubation with the MMP9 antibody. A Cy3 conjugated goat anti-rabbit antibody was used as the secondary antibody. In the image: red staining = MMP9, blue staining = nuclei, green = gelatinase activity.

Western blot - MMP9 antibody - Whole molecule (ab38898)

All lanes : Anti-MMP9 antibody - Whole molecule (ab38898) at 1/1000 dilution

Lane 1 : Human lung tissue lysate at 100 µg with 10% Milk for 1 hour at room temperature
Lane 2 : Human lung tissue lysate at 100 µg with 10% Milk for 1 hour at room temperature
Lane 3 : MMP-9 KO mice tissue lysates with 10% Milk for 1 hour at room temperature
Lane 4 : MMP-9 KO mice tissue lysates with 10% Milk for 1 hour at room temperature

Secondary
HRP-conjugated donkey anti-rabbit polyclonal at 1/1000 dilution
developed using the ECL technique

Performed under reducing conditions.

Specific observed bands 95-100 kDa

This image is courtesy of an anonymous Abreview

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - MMP9 antibody - Whole molecule (ab38898)

ab38898 staining MMP9 in Mouse Pancreatic carcinoma tissue sections by IHC-P (formaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with formaldehyde and blocked with 1% BSA for 1 hour at room temperature. Antigen retrieval was by heat mediation in citric acid (pH6). Samples were incubated with primary antibody (1/100) in 1% Aurion BSA for 12 hours. An HRP-conjugated Donkey polyclonal to rabbit IgG (1/100) was used as secondary antibody.

This image is courtesy of an anonymous Abreview