For the best experience on the Abcam website please upgrade to a modern browser such as Google Chrome
Full length protein corresponding to Mouse MMP9.
Our Abpromise guarantee covers the use of ab38898 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|IHC-P||1/100 - 1/1000. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.|
(see Abreview submitted by Greg Gibson)
We recommend using Goat Anti-Rabbit IgG H&L (Cy3 ®) preadsorbed (ab6939) secondary antibody
|WB||1/1000. Detects a band of approximately 92 kDa. When using colorimetric substrates such as BCIP/NBT use at a 1/5000 dilution (for chemiluminescent substrates). Detects a band of approximately 92-95 kDa (pro-form) and 82kDa (active form) (Human samples). Mouse MMP9 is larger, and on SDS PAGE gels runs about 102-105 kDa. Dilution optimised using Chromogenic detection.|
|IP||Use at an assay dependent concentration.|
|ELISA||Use at an assay dependent concentration.|
|ICC||Use at an assay dependent concentration.|
|IHC-FoFr||Use at an assay dependent concentration. PubMed: 19295156|
This blot was produced using a 4-12% Bis-Tris gel under the MOPS buffer system. The gel was run at 200V for 60 minutes before being transferred onto a nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour before being incubated with anti-MMP9 antibody (ab38898; 2 microgram per mL) overnight at 4°C. Antibody binding was detected using infrared labelled goat anti-rabbit (green) antibody (diluted 1:20000) for 1 hour at room temperature before imaging.
ab38898 detects recombinant Human MMP9 running at ~85 kDa, and endogenous full-length MMP9 in LPS-stimulated cells at ~100 kDa. This antibody also detects a band at 90 kDa in U937 PMA-treated cells.
ab38898 was incubated at 5 ug/mL and ab8245 (loading control to GAPDH) was diluted at 0.1 ug/mL and both were incubated for 48 hours at 4°C. Blots were developed with goat anti-rabbit IgG (H + L) and goat anti-mouse IgG (H + L) secondary antibodies at 1/10 000 dilution for 1 h at room temperature before imaging.
ab38898 staining MMP9 (red) in Mouse Neutrophils and Monocytes cells by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with paraformaldehyde and permeabilized with 2%BSA + 0.2% tritonX100 in PBS. Samples were incubated with primary antibody (1/200 in 2%BSA + 0.2% tritonX100 in PBS) for 25 minutes at 23°C. An Alexa Fluor® 568-conjugated Donkey anti-rabbit IgG polyclonal (1/1000) was used as the secondary antibody. DAPI is stained blue
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. Goat Anti-Rabbit IgG H&L (Cy3 ®) preadsorbed (ab6939) was used as the secondary antibody. In the image: red staining = MMP9, blue staining = nuclei, green = gelatinase activity.
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"