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Human MMP-8(Matrix Metalloproteinase 8) ELISA Kit

Citations(9)Uniprot : P22894

Cat.No.:E-EL-H1450
Reactivity: Human

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Product Details

Properties

Assay type	Sandwich-ELISA
Format	96T/48T
Assay time	3.5h
Detection range	0.16-10 ng/mL
Sensitivity	0.10 ng/mL
Sample type &Sample volume	serum, plasma and other biological fluids; 100μL
Specificity	This kit recognizes Human MMP-8 in samples. No significant cross-reactivity or interference between Human MMP-8 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-8 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-8. 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-8 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-8, 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-8. You can calculate the concentration of Human MMP-8 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-8 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-8 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 (ng/mL)	0.50	1.20	4.40	0.50	1.20	4.10
Standard deviation	0.03	0.06	0.23	0.03	0.06	0.17
CV (%)	6.00	5.00	5.23	6.00	5.00	4.15

Recovery

The recovery of Human MMP-8 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)	86-96	91
EDTA plasma (n=8)	89-102	96
Cell culture media (n=8)	95-111	101

Linearity

Samples were spiked with high concentrations of Human MMP-8 and diluted with Reference Standard & Sample Diluent to produce samples with values within the range of the assay.

Target Information

Database Links	SwissProt: P22894
Synonyms	MMP8, CLG1, HNC, PMNL-CL, Neutrophil Collagenase
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

JOURNAL OF CELLULAR AND MOLECULAR MEDICINE (2021) IF: 5.31
Identification of key candidate biomarkers for severe influenza infection by integrated bioinformatical analysis and initial clinical validation

DOI: 10.1111/jcmm.16275

PMID: 33448094
Sample: Plasma
Journal of Fungi (2021) IF: 5.816
Chronic Pulmonary Aspergillosis: Disease Severity Using Image Analysis and Correlation with Systemic Proinflammation and Predictors of Clinical Outcome

DOI: 10.3390/jof7100842

Sample: peripheral blood
Frontiers in Bioscience-Landmark (2021) IF: 4.009
Association between isotretinoin (Roaccutanne) use and changes in periodontal clinical parameters and MMP-8 and MMP-9 salivary levels.

DOI: 10.52586/4933

Sample: saliva
International Journal of Environmental Research and Public Health (2021) IF: 3.39
Salivary Metalloproteinase-8 and Metalloproteinase-9 Evaluation in Patients Undergoing Fixed Orthodontic Treatment before and after Periodontal Therapy

DOI: 10.3390/ijerph18041583

PMID: 33567492
Sample: Saliva
CLINICAL CHEMISTRY AND LABORATORY MEDICINE (2017) IF: 3.432
Obesity, Metabolic syndrome, Diabetes

DOI: 10.1515/cclm-2017-5030

Sample: serum
BMC Oral Health (2021) IF: 2.757
The effect of conventional versus electronic cigarette use on treatment outcomes of peri-implant disease

DOI: 10.1186/s12903-021-01784-w

Sample: Saliva
JOURNAL OF PERIODONTOLOGY (2019) IF: 2.768
Gingival crevicular fluid semaphorin 4D and peptidylarginine deiminase‐2 levels in periodontal health and disease

DOI: 10.1002/JPER.18-0608

Sample: Gingival crevicular fluid(GCF)
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
Clinical and Experimental Dental Research (2022)
Assessing salivary matrix metalloproteinase-8 in prostate cancer patients undergoing androgen deprivation therapy

DOI: 10.1002/cre2.624

PMID: 35769040
Sample: Salivary

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Reviews/Q&A

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Reviews
Q&A

M********nSubmitted [ Jun 07 2022 ]

Asked: Hello, Does this kit estimated total MMP-8 or active MMP-8 levels ?

adminSubmitted [ Jun 07 2022 ]

Answered: This kit detects total MMP-8.

M********nSubmitted [ Feb 11 2022 ]

Asked: Does this required dilution ?

adminSubmitted [ Feb 11 2022 ]

Answered: Hi Maaz, thanks for your question. Yes, this kit requires dilution, and it's recommended to test normal human serum/plasma samples at about 50-200 fold from our in-house data.

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