Mmp 2

Manufactured by BioLegend

Sourced in United States

MMP-2, also known as Gelatinase A, is a matrix metalloproteinase (MMP) enzyme that plays a role in the breakdown and remodeling of the extracellular matrix. It is responsible for the degradation of type IV collagen, a major component of basement membranes.

Automatically generated - may contain errors

Lab products found in correlation

5 protocols using mmp 2

Proteomic Analysis of ST09 Treatment

Cited 1 time

Check if the same lab product or an alternative is used in the 5 most similar protocols

A total of 75,000 cells/mL were seeded and treated with ST09 (20, 40, 60, and 80 nM) for 48 h and the whole cell lysate was prepared as described [13 (link)]. Next, 30 µg of cell lysates was electrophoresed on 10 to 12% of SDS-PAGE (poly acrylamide gel electrophoresis) and were transferred to a polyvinylidene fluoride membrane (Millipore, Burlington, MA, United States). Blocking was performed using 5% skim milk in 1× PBS and then probed with primary antibodies: MMP2 from Biolegend, MMP1 from elabscience, Apaf, Bad, Bcl2, cytochrome c, Tubulin from Santa-Cruz Biotechnology, CA, and Caspase 9, Caspase 3, PARP, Vimentin, Bax, and GAPDH from Cell Signaling Technology, Beverly, MA, USA, followed by HRP-conjugated secondary anti-rabbit, anti-mouse antibodies (Cell Signaling Technology). The blots were developed using chemiluminescence reagent (Clarity Western ECL blotting substrate, Biorad) and the blot images were captured by the Chemidoc-XRS Biorad gel doc system. The protein band images were quantified using GelQuant.Net, BiochemLab solutions.

Nirgude S., Mahadeva R., Koroth J., Kumar S., Kumar K.S., Gopalakrishnan V., S Karki S, & Choudhary B. (2020). ST09, A Novel Curcumin Derivative, Blocks Cell Migration by Inhibiting Matrix Metalloproteases in Breast Cancer Cells and Inhibits Tumor Progression in EAC Mouse Tumor Models. Molecules, 25(19), 4499.

+ Open protocol

+ Expand

MMP-Mediated Collagen I Degradation Assay

Cited 2 times

Check if the same lab product or an alternative is used in the 5 most similar protocols

MMP degradation assay of extracted type I collagen (Col1) homotrimers or heterotrimers was conducted according to previous description with minor modifications (Han et al., 2010 (link); Makareeva et al., 2010 (link)). As described above, mouse Col1 heterotrimers and homotrimers were extracted from cell culture media of mouse primary pancreatic cancer cell lines and mouse fibroblast line, respectively. MMP-1 (901-MP, R&D Systems) and MMP-2 (Biolegend) were activated before the assay using p-aminophenylmercuric acetate (APMA, A9563, Sigma-Aldrich) according to manufacturer instructions. Col1 samples were prepared in TCNB assay buffer (pH 7.5, 50 mM Tris-HCl, 0.15 M NaCl, 10 mM CaCl₂, 0.05% Brij 35 and incubated with activated MMP-1 (2 nM) or MMP-2 (50 nM) at 37 °C for 20 min. The degradation reaction was terminated by adding 6x reducing SDS Laemmli Sample Buffer (Bio-world) and denatured at 60 °C for 20 min. The degradation of Col1 were examined using Western blotting method showing the cleavage fragments of Col1α1 and Col1α2 chains.

Chen Y., Yang S., Tavormina J., Tampe D., Zeisberg M., Wang H., Mahadevan K., Wu C.J., Sugimoto H., Chang C.C., Jenq R.R., McAndrews K.M, & Kalluri R. (2022). An oncogenic collagen I homotrimer from cancer cells binds to α3β1 integrin and impacts tumor microbiome and immunity to promote pancreatic cancer. Cancer cell, 40(8), 818-834.e9.

+ Open protocol

+ Expand

Zymography Reveals MMP Activity

Check if the same lab product or an alternative is used in the 5 most similar protocols

The 10% zymogram-ready gels (Biorad Laboratories GmbH, Munich, Germany) were loaded with 40 µg protein/probe mixed with a zymography buffer. Recombinant mouse matrix metalloproteinase (MMP)-2 and MMP-9 standards (Biolegend Inc., San Diego, CA) were run as controls (5 ng each). Electrophoresis was run at 200V for 1 hour. After sample separation, gels were incubated twice for 30 minutes in a renaturing buffer and then twice in a denaturing buffer (30 minutes and overnight; Biorad Laboratories GmbH). After washing 3 times for 15 minutes in water, gels were stained for 60 minutes in Simply Blue Safe Stain. Finally, excess dye was removed by washing in water. Clear, ie, nonstained areas, indicated MMP-digested zones.

Witte M., Reiner J., Bannert K., Jaster R., Maschmeier C., Schafmayer C., Lamprecht G, & Berlin P. (2021). Ileocolonic Healing After Extended Small Bowel Resection in Mice: NOD2 Deficiency Impairs Anastomotic Healing and Postoperative Outcome. Inflammatory Bowel Diseases, 27(9), 1503-1512.

+ Open protocol

+ Expand

Characterizing Stem Cell Surface Markers

Check if the same lab product or an alternative is used in the 5 most similar protocols

hWJSCs and hBMMSCs were cultured in a Lab-Tek® Chambered #1.0 Borosilicate coverglass system (Thermo Scientific) until confluence. The cells were then fixed in 4% paraformaldehyde for 10 min, permeabilized with 0.1% Triton-X100 for 10 min, and then washed with PBS. Non-specific blocking was carried out with 10% normal goat serum for 10 min. The cells were incubated with primary antibodies CD29, CD44, CD146, VCAM-1, MMP-2, MMP-9, SDF-1a, ICAM-1, fibronectin, laminin, hyaluronic acid, and collagen type IV (Biolegend, San Diego, USA) at 4 °C overnight. The cells were then incubated with appropriate secondary antibodies (Invitrogen Life Technologies) for 30 min at room temperature in the presence of 1 mg/mL Hoechst 33342 and mounted on to slides with appropriate mounting medium. Photographs were taken using an Olympus FluoView FV1000 laser scanning confocal microscope (Olympus, Japan).

Lin H.D., Fong C.Y., Biswas A, & Bongso A. (2020). Allogeneic human umbilical cord Wharton’s jelly stem cells increase several-fold the expansion of human cord blood CD34+ cells both in vitro and in vivo. Stem Cell Research & Therapy, 11, 527.

+ Open protocol

+ Expand

Profiling Pleural Effusion Biomarkers

Check if the same lab product or an alternative is used in the 5 most similar protocols

The levels of various soluble factors in the pleural effusion were measured using enzyme-linked immunosorbent assay (ELISA) kits for human amphiregulin (R&D Systems), Fibronectin (R&D Systems), MMP-2 (BioLegend, USA), and Macrophage migration inhibitory factor (MIF) (R&D Systems). Other factors, including CCL3, CXCL2, CXCL8, Galectin-3, Galectin-9, amphiregulin, HB-EGF, Intracellular adhesion molecule 1 (ICAM-1), IL-1β, Resistin, Plasminogen activator inhibitor 1 (PAI-1), Tenascin C (TNC), and Vascular endothelial growth factor A (VEGFA) were assessed using a Luminex assay (R&D Systems). The levels of complement system were assessed using human complement panel 1 and 2 bead-based Multiplex Assay kits (EMD Millipore).

Wu Y.Y., Hsu Y.L., Huang Y.C., Su Y.C., Wu K.L., Chang C.Y., Ong C.T., Lai J.C., Shen T.Y., Lee T.H., Hung J.Y, & Tsai Y.M. (2023). Characterization of the pleural microenvironment niche and cancer transition using single-cell RNA sequencing in EGFR-mutated lung cancer. Theranostics, 13(13), 4412-4429.

+ Open protocol

+ Expand

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Revolutionizing how scientists
search and build protocols!