MMP Activity Assay Kit (Fluorometric - Green)

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Lab

Functional Studies - MMP Activity Assay Kit (Fluorometric - Green) (AB112146)

The tests show the MMP activities in different matrices. 25 uL (0.31 ug) APMA-activated MMPs were spiked into 25 ul (6.25e4 cells) of cell lysate. Cells (1e6-5e7 cells) were sonicated in 0.3 mL RIPA buffer with protease inhibitors. Protein amount was determined by BCA method.

Fluorescence shown after subtraction of vehicle control (duplicates, +/- SD).

Purified MMPs used :

MMP1 – ab134442

MMP2 – ab125181

MMP3 – ab96555

MMP8 – ab168050

MMP9 – ab157344

MMP13 – ab134452

MMP14 – ab157068

• FuncS

Lab

Functional Studies - MMP Activity Assay Kit (Fluorometric - Green) (AB112146)

The tests show the MMP activities in different matrices. 25 uL (0.31 ug) APMA-activated MMPs were spiked into 25 ul (0.56 mg) of mouse liver tissue lysate. Frozen tissue was homogenised in 1.2 ml RIPA buffer with protease inhibitors with 1 mm glass beads in a bead beater. Protein was determined by BCA method.

Fluorescence shown after subtraction of vehicle control (duplicates, +/- SD).

Purified MMPs used :

MMP1 – ab134442

MMP2 – ab125181

MMP3 – ab96555

MMP8 – ab168050

MMP9 – ab157344

MMP13 – ab134452

MMP14 – ab157068

• FuncS

Lab

Functional Studies - MMP Activity Assay Kit (Fluorometric - Green) (AB112146)

The tests show the MMP activities in different matrices. 25 uL (0.31 ug) APMA-activated MMPs were spiked into 25 ul (6.25e4 cells) of cell lysate. Cells (1e6-5e7 cells) were sonicated in 0.3 mL RIPA buffer with protease inhibitors. Protein amount was determined by BCA method. Fluorescence shown after subtraction of vehicle control (duplicates, +/- SD).

Purified MMPs used :

MMP1 – ab134442

MMP2 – ab125181

MMP3 – ab96555

MMP8 – ab168050

MMP9 – ab157344

MMP13 – ab134452

MMP14 – ab157068

• FuncS

Lab

Functional Studies - MMP Activity Assay Kit (Fluorometric - Green) (AB112146)

The tests show the MMP activities in different matrices. 25 uL (0.31 ug) APMA-activated MMPs were spiked into 25 ul (0.17 mg) of mouse heart tissue lysate. Frozen tissue was homogenised in 1.2 ml RIPA buffer with protease inhibitors with 1 mm glass beads in a bead beater. Protein was determined by BCA method. Fluorescence shown after subtraction of vehicle control (duplicates, +/- SD).

Purified MMPs used :

MMP1 – ab134442

MMP2 – ab125181

MMP3 – ab96555

MMP8 – ab168050

MMP9 – ab157344

MMP13 – ab134452

MMP14 – ab157068

• FuncS

Lab

Functional Studies - MMP Activity Assay Kit (Fluorometric - Green) (AB112146)

The tests show the MMP activities in different matrices. 25 uL (0.31 ug) APMA-activated MMPs were spiked into 25 ul (0.80 mg) of mouse brain tissue lysate. Frozen tissue was homogenised in 1.2 ml RIPA buffer with protease inhibitors with 1 mm glass beads in a bead beater. Protein was determined by BCA method. Fluorescence shown after subtraction of vehicle control (duplicates, +/- SD).

Purified MMPs used :

MMP1 – ab134442

MMP2 – ab125181

MMP3 – ab96555

MMP8 – ab168050

MMP9 – ab157344

MMP13 – ab134452

MMP14 – ab157068

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Supplier Data

Functional Studies - MMP Activity Assay Kit (Fluorometric - Green) (AB112146)

Detection of activity of MMPs using ab112146.

APMA-activated purified MMPs (30 ng each) were mixed with Green substrate. The fluorescence signal was monitored after 1 hour by using a microplate reader at Ex/Em = 490/525 nm. The reading from all wells was subtracted with the reading from substrate control. Although different MMPs showed different cleavage rate on this MMP substrate, the MMP Green substrate can detect the activity of sub-nanogram of all MMPs (n=3).
NOTE : distinct purified MMP enzymes were used in this test. When using cell extracts, the kit will only detect a general MMP activity and it will not differentiate between the different MMPs.

• FuncS

Lab

Functional Studies - MMP Activity Assay Kit (Fluorometric - Green) (AB112146)

The tests show the MMP activities in different matrices. 25 uL (0.31 ug) APMA-activated MMPs were spiked into 25 ul (0.21 mg) of mouse leg muscle tissue lysate. Frozen tissue was homogenised in 1.2 ml RIPA buffer with protease inhibitors with 1 mm glass beads in a bead beater. Protein was determined by BCA method. Fluorescence shown after subtraction of vehicle control (duplicates, +/- SD).

Purified MMPs used :

MMP1 – ab134442

MMP2 – ab125181

MMP3 – ab96555

MMP8 – ab168050

MMP9 – ab157344

MMP13 – ab134452

MMP14 – ab157068

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PubMed

Functional Studies - MMP Activity Assay Kit (Fluorometric - Green) (AB112146)

Khajah et al investigates endocrine resistant breast cancer cells in response to changes in extracellular pH. MMP activity of pII cells was determined using ab112146. Cells were cultured in a gassed or ungasssed incubator for 1 hour and treated with EGF at 50ng/ml for 30 minutes. MMP activity was then determined using fluorogenic substrate. Fluorescence was measured for excitation/emission of 490/525 nm.

Khajah MA et al., PLoS One 8(10), Fig 8e. doi: 10.1371/journal.pone.0076327. Reproduced under the Creative Commons license http://creativecommons.org/licenses/by/4.0/