MMP 9 ELISA Kit (Matrix Metallopeptidase 9 (Gelatinase B, 92kDa Gelatinase, 92kDa Type IV Collagenase))

Details for Product MMP9 ELISA Kit No. ABIN2506950, Supplier: Log in to see
Antigen
  • CLG4B
  • GELB
  • MANDP2
  • MMP-9
  • AW743869
  • B/MMP9
  • Clg4b
  • pro-MMP-9
  • mmp-9
  • fgMMP-9
  • mmp9
  • ZFMMP-9
  • wu:fb02g06
  • wu:fb07b05
  • wu:fi98c09
  • wu:fj05a08
  • zgc:64165
  • clg4b
  • gelb
  • mandp2
  • matrix metallopeptidase 9
  • collagenase
  • matrix metalloproteinase 9
  • matrix metallopeptidase 9 S homeolog
  • MMP9
  • RB3913
  • BPSS0666
  • Sbal_3732
  • Bcer98_0486
  • Shew185_0630
  • Shal_3056
  • Sbal195_0657
  • Lbys_3550
  • Palpr_2084
  • Mmp9
  • mmp9
  • mmp9.S
Reactivity
Human
Alternatives
Kits with alternative reactivity to:
48
31
17
12
12
10
9
8
4
3
2
1
1
1
1
Method Type
Sandwich ELISA
Detection Range
65.5-16000 pg/mL
Minimum Detection Limit
65.5 pg/mL
Application
ELISA
Options
Supplier
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Supplier Product No.
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Purpose The kit is a sandwich enzyme immunoassay for the in vitro quantitative measurement of MMP-9 in human serum, plasma, and other biological fluids not being tested
Brand EasyTest™
Sample Type Serum, Plasma
Analytical Method Quantitative
Detection Method Colorimetric
Specificity This assay has high sensitivity and excellent specificity for detection of human MMP-9
Sensitivity 22 pg/mL
Characteristics EasyTest ELISA, a fast ELISA provides a simple and rapid quantitation of antigen in a range of sample matrices. Compared to standard ELISA, EasyTest ELISA requires only one wash, decreasing variations and significantly saves time. The whole process requires less than 2.5 hours and provides customers with high accuracy.
Components
  • Assay plate (12x8 coated Microwells)-1
  • Standard-2x 10μL
  • human MMP-9 Mix-4x 18 μL
  • Assay diluent (5x concentrate)-1 x15mL
  • Wash buffer (20x concentrate)-1x25 mL
  • TMB substrate-1x12mL
  • Stop solution-1x 8mL
  • Adhensive strip-2
  • Instruction manual-1.
Material not included 1. Distilled or deionized water, 2.Precision pipettes, with disposable plastic tips, 3.Beakers, flasks, cylinders necessary for preparation of reagents, 4.Microplate washing device (multichannel pipette or automated microplate washer), 5.Microplate shaker, 6.Microplate reader capable of reading at 450 nm.
Alternative Name MMP-9 (MMP9 ELISA Kit Abstract)
Background Matrix metalloproteinases (MMPs) are a family of zinc-dependent endopeptidases that degrade extracellular matrix proteins.MMP-9 is involved in inflammation, tissue remodeling, wound healing, mobilization of matrix bound growth factors and processing of cytokines.
Molecular Weight 92 kDa
Gene ID 4318
NCBI Accession NP_004985
UniProt P14780
Research Area Extracellular Matrix
Pathways Cellular Response to Molecule of Bacterial Origin, Positive Regulation of Immune Effector Process, CXCR4-mediated Signaling Events
Application Notes Optimal working dilution should be determined by the investigator.
Sample Volume 50 μL
Assay Time 1.5 h
Plate Pre-coated
Protocol This assay employs the fast quantitative sandwich enzyme immunoassay technique. Antibody specific for MMP-9 has been pre-coated onto a microplate. Standards, samples and human MMP-9 mix are pipetted into the wells and any MMP-9 present is bound by the immobilized antibody. Following a wash to remove any unbound reagent, a TMB substrate solution is added to the wells and color develops in proportion to the amount of MMP-9 bound in the initial step. The color development is stopped and the intensity of the color is measured.
Reagent Preparation
  1. Assay diluent: Dilute the concentrated assay diluent 1:5 with distilled water (e.g. 10 mL plus 40 mL). 2. Wash buffer: Dilute the concentrated wash buffer 1:20 with distilled water (e.g. 20 mL plus 380 mL). 3. Standard: Briefly spin standard vial before use. Add 390 μL 1x Assay Diluent to prepare a 50 ng/mL standard. Gently vortex to mix.Take 160 μL MMP-9 standard into a tube, then add 340 μL 1x Assay Diluent to prepare a 16000 pg/mL stock standard solution.Add 150 μL 1x Assay Diluent to 7 tubes. Perform serial dilutions by adding 100 μL of each standard to the next tube and vortexing between each transfer, the last tube with 1x assay diluent is the blank as 0pg/mL. 4. human MMP-9 Mix: Within 15 minutes prior to use, briefly spin the vial. Add 1482 μL of 1x Assay diluent to the vial and mix by pipetting. A vial mix can be used for about 30 wells.
Sample Collection Serum: Use a serum separator tube (SST) and allow samples to clot for two hours at room temperature or overnight at 4 °C before centrifugation for 15 minutes at 1000 x g. Remove serum and assay immediately or aliquot and store samples at -20 °C or -80 °C. Avoid repeated freeze-thaw cycles.
Plasma: Collect plasma using EDTA, or citrate or heparin as an anticoagulant. Centrifuge for 15 minutes at 1000 x g at 2-8 °C within 30 minutes of collection. Assay immediately or aliquot and store samples at -20 °C or -80 °C. Avoid repeated freeze-thaw cycles.
Sample Preparation

Levels of the target protein may vary among different specimens. Optimal dilution factors for each sample must be determined by the investigator,The dilution scheme is only suggestion: the recommended dilution for serum and plasma is 1:300.

Assay Procedure
  1. All reagents must be brought to room temperature (18-25 °C) prior to use 2. Prepare all reagents, working standards,samples and mix as directed in the respective sections. 3.Determine the number of wells to be used and put any remaining wells and the desiccant back into the pouch and seal the ziploc, store unused wells at 4 °C. 4. Add 50μL of standard or sample to the designated wells, Gentlely shake/tap the plate for 5 second to mix. 5. Add 50 μL of MMP-9 Mix into all wells, including the blank wells, cover the plate with adhensive strip. Incubate for 1 hour at room temperature with gentle shaking. 6.Decant or aspirate contents of wells. Wash wells by filling with at least 300 μL/well prepared wash buffer followed by decanting/aspirating. Soak wells in wash buffer for 30 seconds to 1 minute for each wash. Repeat wash 4 times for a total of 5 washes. After the last wash, blot plate on absorbent paper to remove residual buffer. Thorough washing at this step is very important, complete removal of liquid is required for proper performance. 7.Pipette 100 μL of TMB Substrate Solution to each well. Incubate plate for 15 minutes at room temperature in the dark with gentle shaking. 8. Add 50μL of Stop Solution to each well, gently tap the plate to ensure thorough mixing. 9. Determine the optical density of each well within 15 minutes using a microplate reader set to 450nm. If wavelength correction is available, set to 570nm. Subtract readings at 570nm from the readings at 450nm. This subtraction will correct for optical imperfections in the plate.
Calculation of Results
  1. Average the duplicate readings for each standard and sample and subtract the average zero standard optical density.2. Create a standard curve by reducing the data using computer software capable of generating a four parameter logistic (4-PL) curve fit. As an alternative, construct a standard curve by plotting the mean absorbance for each standard on the x-axis against the concentration on the y-axis and draw a best fit curve through the points on the graph. The data may be linearized by plotting the log of the target antigen concentration versus the log of the O.D. and the best fit line can be determined by regression analysis. This procedure will produce an adequate but less precise fit of the data. 3. If samples have been diluted, the concentration read from the standard curve must be multiplied by the dilution factor.
Assay Precision intra CV<10%, inter CV <15%
Restrictions For Research Use only
Buffer 0.05 % Proclin 300
Preservative ProClin
Precaution of Use The Stop Solution provided with this kit is an acid solution. Wear eye, hand, face and clothing protection when using this material.
Storage 4 °C/-20 °C
Supplier Images
ELISA image for Matrix Metallopeptidase 9 (Gelatinase B, 92kDa Gelatinase, 92kDa Type IV Collagenase) (MMP9) ELISA Kit (ABIN2506950) Matrix Metallopeptidase 9 (Gelatinase B, 92kDa Gelatinase, 92kDa Type IV Collagenase) (MMP9) ELISA Kit
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