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 (Collagenase IV) Rabbit Polyclonal Antibody (PA516509) at 2ug/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 conjugate (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 (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|
|Published species reactivity||Human|
|Host / Isotype||Rabbit / IgG|
|Immunogen||A synthetic peptide from the middle region of human MMP-9|
|Storage buffer||PBS, pH 7.4, with 0.2% BSA|
|Contains||0.09% sodium azide|
|Storage Conditions||4° C|
|Tested Applications||Dilution *|
|Flow Cytometry (Flow)||3-5 µg/10^6 cells|
|Immunocytochemistry (ICC)||2 µg/ml|
|Immunofluorescence (IF)||2 µg/ml|
|Immunohistochemistry (Paraffin) (IHC (P))||1:20-1:100|
|Western Blot (WB)||2-4 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.
|Immunohistochemistry (IHC)||See 2 publications below|
PA5-16509 targets MMP-9 (92kDa Collagenase IV) in WB applications and shows reactivity with Guinea Pig and Human samples.
The PA5-16509 immunogen is a synthetic peptide from the middle region of human MMP-9.
MMPs are a group of enzymes involved in matrix degradation. They share some important characteristics, such as a common mode of activation, a conserved amino acid sequence in the putative metal binding-active site region, and inhibition by specific proteinase inhibitors known as tissue inhibitors of metalloproteinases (TIMPs). Reportedly, MMPs play a crucial role in tumor cell invasion and metastasis.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
Matrix metalloproteinases 2 and 9 and e-cadherin expression in the endometrium during the implantation window of infertile women before in vitro fertilization treatment.
PA5-16509 was used in immunohistochemistry to study the implantation window expression of endometrial MMP-2, MMP-9 and E-cadherin in infertile woman prior to the initiation of IVF treatment
|Maia-Filho VO,Rocha AM,Ferreira FP,Bonetti TC,Serafini P,Motta EL||Reproductive sciences (Thousand Oaks, Calif.) (22:416)||2015|
Effect of the expression of matrix metalloproteases and their tissue inhibitors on survival of patients with resectable colorectal cancer.
PA5-16509 was used in immunohistochemistry to study the expression and clinical relevance of MMPs and TIMPs in patients with resectable colorectal carcinoma
|González L,Eiró N,González LO,Andicoechea A,Barbón E,García-Muñiz JL,Vizoso FJ||Digestive diseases and sciences (57:2063)||2012|