Western blot analysis was performed on whole cell extracts (30 µg lysate) of HT-1080 (Lane 1), MDA-MB-231 (Lane 2) and A549 (Lane 3). The blot was probed with Rabbit Anti-MMP14 Polyclonal Antibody (Product # PA5-16514, 2µg/ml) and detected by chemiluminescence using Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 0.4 µg/ml, 1:2500 dilution). A 65 kDa band corresponding to MMP14 was observed across the cell lines tested, an additional band of 160 kDa corresponding to the oligomer form of MMP-14 was also detected. Known quantity of protein samples were electrophoresed using Novex® NuPAGE® 4-12 % Bis-Tris gel (Product # NP0321BOX), XCell SureLock™ Electrophoresis System (Product # EI0002) and Novex® Sharp Pre-Stained Protein Standard (Product # LC5800). Resolved proteins were then transferred onto a nitrocellulose membrane with iBlot® 2 Dry Blotting System (Product # IB21001). The membrane was probed with the relevant primary and secondary Antibody following blocking with 5 % skimmed milk. Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005).
|Tested species reactivity||Human, Mouse|
|Host / Isotype||Rabbit / IgG|
|Immunogen||A synthetic peptide from the hemopexin-like domain of human MMP-14 / MT1-MMP|
|Storage buffer||PBS, pH 7.4, with 0.2% BSA|
|Contains||0.09% sodium azide|
|Storage Conditions||4° C|
|Tested Applications||Dilution *|
|Western Blot (WB)||2 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.
PA5-16514 targets MMP-14 in WB applications and shows reactivity with mouse and Human samples.
The PA5-16514 immunogen is a synthetic peptide from the hemopexin-like domain of human MMP-14 / MT1-MMP.
MMPs are a group of enzymes involved in matrix degradation. Of the sixteen members of matrix metalloproteinase family ten exist in soluble form whereas MT-MMPs exist as integral membrane proteins. MT1-MMP, MT2-MMP, MT3-MMP also known as MMP14, MMP15, MMP16 respectively contain a C-terminal transmembrane domain anchoring them to the cell surface. They have an 8 amino acid insert in their catalytic domain. MT1-MMP cleaves progelatinase A (MMP-2) and progelatinase B to their active forms. MT1-MMP gets activated through the membrane plasmin cascade. It binds to TIMP-2 first and then to MMP-2 forming a trimer. It cleaves the pro-MMP-2 to its active form.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.