Immunofluorescence analysis of MMP-9 was performed using 70% confluent log phase MCF-7 cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 10 minutes, and blocked with 1% BSA for 1 hour at room temperature. The cells were labeled with MMP-9 (92kDa Collagenase IV) Rabbit Polyclonal Antibody (Product # PA5-16851) at 2µg/ml in 0.1% BSA and incubated for 3 hours at room temperature and then labeled with Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjµgate (Product # A27034) at a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (Product # S36938). F-actin (Panel c: red) was stained with Alexa Fluor® 555 Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing cytoplasmic localization. Panel e shows the no primary antibody control. The images were captured at 60X magnification.
|Tested species reactivity||Guinea pig, Human|
|Host / Isotype||Rabbit / IgG|
|Immunogen||A synthetic peptide derived from near C-terminal of human MMP-9 protein|
|Storage buffer||PBS, pH 7.6, with 0.2% BSA|
|Contains||15mM sodium azide|
|Storage Conditions||4° C|
|Tested Applications||Dilution *|
|Immunocytochemistry (ICC)||2 µg/ml|
|Immunofluorescence (IF)||2 µg/ml|
|Immunohistochemistry (Paraffin) (IHC (P))||1:100|
|Western Blot (WB)||1-3 µg/ml|
* Suggested working dilutions are given as a guide only. It is recommended that the user titrate the product for use in their own experiment using appropriate negative and positive controls.
PA5-16851 targets MMP-9 (92kDa Collagenase IV) in IHC (P) and WB applications and shows reactivity with Guinea Pig and Human samples.
The PA5-16851 immunogen is a synthetic peptide derived from near C-terminal of human MMP-9 protein.
Metallomatrix proteases (MMPs) are proteolytic enzymes capable of degrading connective tissue components. They have a common mode of activation, a conserved amino acid sequence in the putative metal binding-active site region, and are inhibited by specific tissue inhibitors of metalloproteinases (TIMPs). MMPs and TIMPs play a significant role in regulating angiogenesis. This antibody reacts with both latent and active MMP-9.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.