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Human MMP-13(Matrix Metalloproteinase 13) ELISA Kit

Citations(20)Uniprot : P45452

Cat.No.:E-EL-H6023
Reactivity: Human

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Product Details

Properties

Assay type	Sandwich-ELISA
Format	96T/48T
Assay time	3.5h
Detection range	15.63-1000 pg/mL
Sensitivity	9.38 pg/mL
Sample type &Sample volume	serum, plasma and other biological fluids; 100μL
Specificity	This kit recognizes Human MMP-13 in samples. No significant cross-reactivity or interference between Human MMP-13 and analogues was observed.
Reproducibility	Both intra-CV and inter-CV are < 10%.
Application	This ELISA kit applies to the in vitro quantitative determination of Human MMP-13 concentrations in serum, plasma and other biological fluids.

Test Principle

This ELISA kit uses the Sandwich-ELISA principle. The micro ELISA plate provided in this kit has been pre-coated with an antibody specific to Human MMP-13. Standards or samples are added to the micro ELISA plate wells and combined with the specific antibody. Then a biotinylated detection antibody specific for Human MMP-13 and Avidin-Horseradish Peroxidase (HRP) conjugate are added successively to each micro plate well and incubated. Free components are washed away. The substrate solution is added to each well. Only those wells that contain Human MMP-13, biotinylated detection antibody and Avidin-HRP conjugate will appear blue in color. The enzyme-substrate reaction is terminated by the addition of stop solution and the color turns yellow. The optical density (OD) is measured spectrophotometrically at a wavelength of 450 nm ± 2 nm. The OD value is proportional to the concentration of Human MMP-13. You can calculate the concentration of Human MMP-13 in the samples by comparing the OD of the samples to the standard curve.

Kit components & Storage

An unopened kit can be stored at 2-8℃ for 1 month. If the kit is not supposed to be used within 1 month, store the components separately according to the following conditions once the kit is received.

Item	Specifications	Storage
Micro ELISA Plate(Dismountable)	96T: 8 wells ×12 strips 48T: 8 wells ×6 strips	-20℃, 6 months
Reference Standard	96T: 2 vials 48T: 1 vial
Concentrated Biotinylated Detection Ab (100×)	96T: 1 vial, 120 μL 48T: 1 vial, 60 μL
Concentrated HRP Conjugate (100×)	96T: 1 vial, 120 μL 48T: 1 vial, 60 μL	-20℃(Protect from light), 6 months
Reference Standard & Sample Diluent	1 vial, 20 mL	2-8°C, 6 months
Biotinylated Detection Ab Diluent	1 vial, 14 mL
HRP Conjugate Diluent	1 vial, 14 mL
Concentrated Wash Buffer (25×)	1 vial, 30 mL
Substrate Reagent	1 vial, 10 mL	2-8℃(Protect from light)
Stop Solution	1 vial, 10 mL	2-8°C
Plate Sealer	5 pieces
Manual	1 copy
Certificate of Analysis	1 copy

Technical Data

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and high level Human MMP-13 were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level Human MMP-13 were tested on 3 different plates, 20 replicates in each plate.

	Intra-assay Precision			Inter-assay Precision
Sample	1	2	3	1	2	3
n	20	20	20	20	20	20
Mean (pg/mL)	43.41	107.90	450.13	44.04	101.12	413.83
Standard deviation	2.34	4.95	22.82	2.53	5.06	19.66
CV (%)	5.39	4.59	5.07	5.74	5.00	4.75

Recovery

The recovery of Human MMP-13 spiked at three different levels in samples throughout the range of the assay was evaluated in various matrices.

Sample Type	Range (%)	Average Recovery (%)
Serum(n=8)	95-106	98
EDTA plasma (n=8)	98-110	105
Cell culture media (n=8)	90-103	96

Linearity

Samples were spiked with high concentrations of Human MMP-13 and diluted with Reference Standard & Sample Diluent to produce samples with values within the range of the assay.

Target Information

Database Links	SwissProt: P45452
Synonyms	MMP13, CLG3, MANDP1, Collagenase 3
Research Area	Cancer, Cell Biology, Cardiovascular, Signal transduction

Assay Procedures

	1. Add 100μL standard or sample to the wells. Incubate for 90 min at 37°C
	2. Discard the liquid, immediately add 100μL Biotinylated Detection Ab working solution to each well. Incubate for 60 min at 37°C
	3. Aspirate and wash the plate for 3 times
	4. Add 100μL HRP conjugate working solution. Incubate for 30 min at 37°C. Aspirate and wash the plate for 5 times
	5. Add 90μL Substrate Reagent. Incubate for 15 min at 37°C
	6. Add 50μL Stop Solution
	7. Read the plate at 450nm immediately. Calculation of the results

Citations

OSTEOARTHRITIS AND CARTILAGE (2018) IF: 5.454
Associations between serum S100A8/S100A9 and knee symptoms, joint structures and cartilage enzymes in patients with knee osteoarthritis

DOI: 10.1016/j.joca.2018.08.020

PMID: 30240939
Sample: Serum
Frontiers in Pharmacology (2021) IF: 5.811
Single Cell RNA-Seq Analysis Identifies Differentially Expressed Genes of Treg Cell in Early Treatment-Naive Rheumatoid Arthritis By Arsenic Trioxide

DOI: 10.3389/fphar.2021.656124

PMID: 34108876
Sample: Cell sample
Spine Journal (2022) IF: 4.166
Extracellular vesicle and soluble fractions of adipose tissue-derived mesenchymal stem cells secretome induce inflammatory cytokines modulation in an in vitro model of discogenic pain

DOI: 10.1016/j.spinee.2022.01.012

PMID: 35121152
Sample: human adipose-derived mesenchymal stem cells (HAdM
OSTEOARTHRITIS AND CARTILAGE (2017) IF: 4.742
Associations between serum ghrelin and knee symptoms, joint structures and cartilage or bone biomarkers in patients with knee osteoarthritis

DOI: 10.1016/j.joca.2017.05.015

Sample: Serum
INTERNATIONAL IMMUNOPHARMACOLOGY (2019) IF: 4.932
Arsenic trioxide improves Treg and Th17 balance by modulating STAT3 in treatment-naïve rheumatoid arthritis patients

DOI: 10.1016/j.intimp.2019.05.001

Sample: PBMCs cells
PPAR Research (2019) IF: 4.964
Arsenic Trioxide in Synergy with Vitamin D Rescues the Defective VDR-PPAR- Functional Module of Autophagy in Rheumatoid Arthritis

DOI: 10.1155/2019/6403504

Sample: rheumatoid arthritis (RA) fibroblast-like synovioc
Cartilage (2023) IF: 3.117
Ferroptosis Plays a Role in Human Chondrocyte of Osteoarthritis Induced by IL-1β In Vitro

DOI: 10.1177/19476035221142011

Sample: Chondrocyte
Bioengineered (2022) IF: 3.269
The protective effects of Olmesartan against interleukin-29 (IL-29)-induced type 2 collagen degradation in human chondrocytes

DOI: 10.1080/21655979.2021.1997090

PMID: 35012432
Sample: T/C-28a2 cell
BIOMEDICINE & PHARMACOTHERAPY (2018) IF: 3.457
MicroRNA-449a upregulation promotes chondrocyte extracellular matrix degradation in osteoarthritis

DOI: 10.1016/j.biopha.2018.06.074

Sample: Cell culture supernatant
PHYTOMEDICINE (2018) IF: 3.61
Ethanolic extract of Kaempferia parviflora interrupts the mechanisms-associated rheumatoid arthritis in SW982 culture model via p38/STAT1 and STAT3 pathways

DOI: 10.1016/j.phymed.2018.11.015

PMID: 31005814
Sample: Cell culture supernatant
EXPERIMENTAL CELL RESEARCH (2017) IF: 3.905
MicroRNA-132 upregulation promotes matrix degradation in intervertebral disc degeneration

DOI: 10.1016/j.yexcr.2017.08.011

Sample: Cell culture supernatant
CLINICAL RHEUMATOLOGY (2019) IF: 2.293
Associations between serum IL-8 and knee symptoms, joint structures, and cartilage or bone biomarkers in patients with knee osteoarthritis

DOI: 10.1007/s10067-019-04718-8

PMID: 31377918
Sample: Serum
Experimental and Therapeutic Medicine (2021) IF: 2.447
Vicenin 3 ameliorates ECM degradation by regulating the MAPK pathway in SW1353 chondrocytes

DOI: 10.3892/etm.2021.10896

Sample: SW1353 cell
BIOSCIENCE REPORTS (2019) IF: 2.535
Association of IL-6, IL-8, MMP-13 gene polymorphisms with knee osteoarthritis susceptibility in the Chinese Han population

DOI: 10.1042/BSR20181346

PMID: 30635366
Sample: Tissue homogenate
Frontiers in Surgery (2021) IF: 2.718
Potential Value of Matrix Metalloproteinase-13 as a Biomarker for Osteoarthritis

DOI: 10.3389/fsurg.2021.750047

Sample: serum
PLoS One (2020) IF: 2.74
Protective effects of stem cells from human exfoliated deciduous teeth derived conditioned medium on osteoarthritic chondrocytes

DOI: 10.1371/journal.pone.0238449

PMID: 32886713
Sample: Cell culture medium
JOURNAL OF BIOENERGETICS AND BIOMEMBRANES (2022) IF: 2.945
Circ-LRP1B functions as a competing endogenous RNA to regulate proliferation, apoptosis and oxidative stress of LPS-induced human C28/I2 chondrocytes

DOI: 10.1007/s10863-022-09932-9

PMID: 35274224
Sample: C28/I2 cell
EXPERIMENTAL AND CLINICAL ENDOCRINOLOGY & DIABETES (2017) IF: 1.685
Expression and Significance of MMPs in Synovial Fluid, Serum and PBMC Culture Supernatant Stimulated by LPS in Osteoarthritis Patients With or Without Diabetes

DOI: 10.1055/s-0043-122223

PMID: 29270923
Sample: Serum,Synovia
Molecular Medicine Reports (2019) IF: 1.851
Chrysin protects human osteoarthritis chondrocytes by inhibiting inflammatory mediator expression via HMGB1 suppression

DOI: 10.3892/mmr.2018.9724

PMID: 30535473
Sample: Cell culture medium
International Journal of Rheumatic Diseases (2020) IF: 1.98
Associations of serum citrate levels with knee structural changes and cartilage enzymes in patients with knee osteoarthritis

DOI: 10.1111/1756-185X.13787

Sample: serum

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