MMPs-SensoLyte® Assays & Enzymes


AnaSpec’s SensoLyte® MMP assay kits, especially the SensoLyte® 520 MMP Assay Kits, have become the new standard in MMP assay sensitivity. With published results in world-class journals and the industry’s longest wavelength, the SensoLyte® 520 line is one of the most sensitive (Figure 1) and brightest lines of MMP assay kits in the world.

SensoLyte® 520 MMP Assays

Measure MMP activity
96-well or 384-well format
Mix and read
Fluorimetric - long wavelength
5-FAM/QXL™ 520 FRET pair (Ex/Em=490/520 nm)
Minimal autofluorescence
Assay time – as short as 1h

Figure 1. The performance of an identical peptide sequence, one coupled with 5-FAM/QXLTM 520 and the other coupled with EDANS/DABCYL FRET pairs was compared (A). Fluorescence was measured after 1 hr incubation of FRET substrates with serial dilutions of MMP-13 (Flex800, Bio-Tek instruments). A 5-FAM/QXLTM 520 MMP substrate was compared with Mca/Dpa FRET substrate (B). Fluorescence was measured after 1 hr incubation of substrates using serial dilutions of MMP-13 (Flexstation 384II, Molecular Devices).

SensoLyte® Plus 520 MMP Assays

Measure MMP activity
Enhanced specificity – human capture antibody included
96-well format (strips)
Mix and read
Fluorimetric - long wavelength
5-FAM/QXL™ 520 FRET pair (Ex/Em=490/520 nm)
Minimal autofluorescence

SensoLyte® 520 Series

SensoLyte Plus® 520 Series

SensoLyte® 490 Series

SensoLyte® ELISA Series

Recombinant Human MMPs

MMP-1

71150

72012

71128

72102

72004

MMP-2

71151

71129

72005

MMP-3

71152

71130

72103

72006

MMP-7

71153

71132

72007

MMP-8

71154

71133

72104

72008

MMP-9

71155

72017

71134

72105

72009

72069 (mouse)

MMP-10

72024

72106

72067

MMP-12

71157

71137

72010

MMP-13

71156

72019

71135

72107

72011

MMP-14

72025

72068

SensoLyte® 490 MMP Assays

Measure MMP activity
96-well or 384-well format
Mix and read
Fluorimetric
EDANS/DABCYL FRET pair (Ex/Em=340/490 nm), except 71132 NBD/DMC FRET pair (Ex/Em=370/460 nm)
Assay time – as short as 1h

SensoLyte® MMP ELISA Assays

Measure MMP amount
96-well format
Colorimetric (Absorbance)

Also available:
SensoLyte® 520 Generic MMP Assay Kit
*Fluorimetric*

SensoLyte® 520 MMP Substrate Sampler Assay Kit *Fluorimetric*
SensoLyte® 520 MMP Profiling Assay Kit *Fluorimetric*

MMP Recombinant Proteins

100 ul (10 ug/ml)
Highly active after APMA incubation
Serve as positive controls for SensoLyte® MMP assay kits
Can also be used for zymography, Western blot, IP, and ELISA

Anti-MMP Antibodies

Human, zebrafish (Z-Fish™), mouse and rat specific
Epitopes from hinge,C-terminal (CT), N-terminal (NT), IN (internal) regions

Posters Presented at Scientific Meetings:

Table. Publications citing the use of AnaSpec’s SensoLyte® MMP assay kits and their applications.

Cat#

Product

Application

71150

SensoLyte™ 520 MMP-1 Assay Kit

MMP-1 activity measured in hyperhomocysteinemic rats.1

71151

SensoLyte™ 520 MMP-2 Assay Kit

(human) pro-MMP-2 … first activated with 1 mm APMA for 20 min or 2 h…2, 3

…tissue extracts were obtained from nonpregnant, pregnant, and postpartum mice…and a fluorescently labeled substrate specific for MMP2 was used to determine enzyme activity.4

WT, Fbln3–/–, and Fbln5–/– mice were anesthetized and perfused with 10% neutral buffered formalin (pH 7.4). Thereafter, the female urogenital tract was dissected en bloc…Vaginal tissues were thawed on ice, minced, washed in PBS, and homogenized in MMP2 assay buffer (EnzoLyte 520 MMP-2 assay kit; AnaSpec, San Jose, CA)…5

Rat aortic SMCs(smooth muscle cells) and adventitial FBs (fibroblasts) and MFBs (myofibroblasts) were obtained…The MMP-1 and MMP-2 activities in the conditioned media of control and 1-h flow- and 6-h flow-treated gels were determined using the SensoLyte Plus 520 MMP-1 Assay Kit and SensoLyte 520 MMP-2 Assay Kit from AnaSpec (San Jose, CA).6

71155

SensoLyte™ 520 MMP-9 Assay Kit

(human) … pro-MMP-9 … first activated with 1 mm APMA for 20 min or 2 h…2, 3

71156

SensoLyte™ 520 MMP-13 Assay Kit

MMP-13 (rat interstitial collagenase) activity was detected at 0 and 7 days using the SensoLyte™ (formerly Enzolyte) 520 MMP-13 assay kit.7

72017

SensoLyte Plus™ 520 MMP-9 Assay Kit

MMP-9 activities of cell [MiaPaCa2 and PANC1 human pancreatic ductal adenocarcinoma cells]or tumor lysates were assessed using the SensoLyte™ (formerly Enzolyte) Plus 520 Enhanced Selectivity MMP-9 activity.8

… genetically altered cells (esophageal squamous) were used to detect MMP-9 enzymatic activity in duplicate using the SenzoLyte Plus MMP-9 assay kit from AnaSpec.9

Rat primary microglia (5 × 106 cells/mL) were treated with LPS/PMA in the presence or absence of MMP inhibitors for 24 h and the supernatants were collected for measuring MMP activities. Both the total and active MMP-3 or MMP-9 activities were assayed according to SensoLyteTM 520 MMP-3 or SensoLyte PlusTM 520 MMP-9 assay system, respectively (AnaSpec, San jose, CA, USA). 10

72019

SensoLyte Plus™ 520 MMP-13 Assay Kit

MMP-13 activity (in RKO- human colorectal carcinoma cell line) cells using the SensoLyte Plus™ 520 MMP-13 Assay.11

Adult human mesenchymal stem cells were encapsulated in pure PEG hydrogels, without peptide functionality, at a concentration of 2 × 106 cells/ml… Cell/polymer constructs were removed from culture at specific time points and analyzed for production of MMP-13 via the SensoLyte Plus™ (formerly EnzoLyte Plus™) 520 MMP-13 Assay Kit (AnaSpec).12

Serum-free D-MEM (Dulbecco's modified Eagle medium) conditioned by confluent stromal cell cultures for 24 h was concentrated approximately 40 fold…and analyzed using SensoLyte Plus 520 MMP-1 and SensoLyte Plus 520 MMP-13 Assay Kits (AnaSpec; San Jose, California, USA).13

71158

SensoLyte™ 520 Generic MMP Assay Kit

General MMP activity (from rat heart) was measured using the SensoLyte™ (formerly Enzolyte) 520 Generic MMP assay kit.14

General MMP activity was assessed using the EnzoLyte™ 520 Generic MMP assay Kit (AnaSpec, San Jose, CA). This kit is optimized to detect the activity of different MMPs such as MMP-1, -2, -3, -7, -8, -9, -12, -13 and -14, using a fluorescence-quenched substrate (5-FAM/QXL™520).15

The SensoLyte (formerly EnzoLyte) 520 Generic MMP Fluorometric Assay Kit (AnaSpec) was used according to the manufacturer's directions. Cell culture supernatants were incubated with 1 mmol/L of 4-aminophenylmercuric acetate for 24 h at 37°C to activate pro-MMPs. 16

WT mice of the genetic background C57BL/6 were obtained from Jackson Laboratories…MMP activity was determined using the SensoLyte 520 Generic MMP Assay Kit (AnaSpec) as per manufacturer's instructions. Briefly, media from WT, TIMP-3–/–, and WT cells treated with rTIMP-3 (1 µg/ml) were collected 72 h postseeding, and 50 µl of these media were incubated with the FAM/QXL 520 fluorescence resonance energy transfer substrate for 1 h in a black 96-well plate at room temperature in the dark. 17

Thirty human fibroblast populations were isolated from the skin biopsies of SSc(Systemic sclerosis) patients…Total MMP activity was determined in the same supernatant as that for collagen assay using the SensoLyteTM 520 Generic MMP assay kit (Anaspec, San Jose, CA, USA). This kit can detect simultaneously the activity of MMP-1, 2, 7, 8, 9, 12, 13 and 14.18

71170

SensoLyte™ 520 MMP Substrate Sampler Kit

Aspergillus culture supernatants, human serum and the mixture of both..(using)..fluorescence resonance energy transfer (FRET) were chosen out of a commercially available protease assay kit comprising 16 different substrates SensoLyte™ (formerly Enzolyte) 520 matrixmetalloprotease (MMP) substrate sample kit, AnaSpec.19

The activity of MMPs in the conditioned medium and cell lysates (CHO or APP-overexpressed neuroblastoma cells) was determined using SensoLyte™ (formerly EnzoLyte) MMP fluorometric assay kit.20

Murine Metastatic 4T1 and normal NmuMG cells were obtained…The activated samples were assay for MMP-9 activity using the SensoLyte 520 MMP Sampler Kit (AnaSpec).21

72025

SensoLyte® 520 MMP-14 Assay Kit

To evaluate MT1-MMP activity, we prepared membrane fractions of HAECs (human aortic endothelial cells)…cells were lysed with a hypotonic buffer, then sonicated and centrifuged at 15 000 g for 10 min. The separated membrane fractions were assessed by the commercially available fluorescent assay kit (SensoLyte 520 MMP-14 assay kit, AnaSpec, San Jose, CA, USA) according to the manufacturer's instructions. 22

72012

SensoLyte® Plus 520 MMP-1 Assay Kit

Rat aortic SMCs(smooth muscle cells) and adventitial FBs (fibroblasts) and MFBs (myofibroblasts) were obtained…The MMP-1 and MMP-2 activities in the conditioned media of control and 1-h flow- and 6-h flow-treated gels were determined using the SensoLyte Plus 520 MMP-1 Assay Kit and SensoLyte 520 MMP-2 Assay Kit from AnaSpec (San Jose, CA). 6

Primary cell cultures were established from biopsy and resection specimens from patients pre-operatively diagnosed with GCT…Serum-free D-MEM conditioned by confluent stromal cell cultures for 24 h …analyzed using SensoLyte Plus 520 MMP-1 and SensoLyte Plus 520 MMP-13 Assay Kits (AnaSpec; San Jose, California, USA). 23

71152

SensoLyte® 520 MMP-3 Assay Kit

Rat primary microglia (5 × 106 cells/mL) were treated with LPS/PMA in the presence or absence of MMP inhibitors for 24 h and the supernatants were collected for measuring MMP activities. Both the total and active MMP-3 or MMP-9 activities were assayed according to SensoLyteTM 520 MMP-3 or SensoLyte PlusTM 520 MMP-9 assay system, respectively (AnaSpec, San Jose, CA, USA).10

Astrocytes were prepared from the cerebrum of 1–2-day-old Wistar rats…Stromelysin-1 activity in the astrocytic culture medium was determined using a stromelysin-1 (MMP-3) activity assay kit (SensoLyte 520 MMP-3 assay kit, AnaSpec, San Jose, CA).24

60580

520 MMP FRET Substrate XIII

The activity of MMP-3 (stromelysin-1) (of rat brain homogenates) was measured fluorometrically using a 5-FAM/QXL520 FRET peptide…25

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