PromedeusLab MMP-9 ELISA, Human

PL1014

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Description

Enzyme immunoassay for the quantitative determination of matrix metalloproteinase 9 (MMP-9) in human serum and plasma samples. Quality controls are included with the kit. Developed and manufactured in Europe by PromedeusLab.

Summary

Matrix metalloproteinases (MMPs) are zinc-dependent endopeptidases, responsible for the integrity of the basement membrane (BM) via degradation of extracellular matrix and BM components (1). MMPs are presented in various types of cells, including cardiomyocytes, cerebral neurons, hepatocytes, and many others, thus influencing various processes.

Increased matrix metalloprotease-9 (MMP-9), also called Gelatinase B, after myocardial infarction (MI) exacerbates ischemia-induced chronic heart failure (CHF) (2). It is also upregulated in the diabetic heart, and ablation of MMP-9 decreases infarct size in the nondiabetic myocardial infarction heart (3). There is also strong evidence that the increased levels of MMP-9 are associated with many inflammatory conditions, such as coronary artery disease, COPD, arthritis, and metabolic syndrome (4). It was also proved that MMP-9 is associated with progression of atherosclerosis and higher risk of cardiovascular events. It was also suggested that elevated levels of MMP-9 (combined with MMP-2) are associated with highly tumorigenic cancers.

Other Names

Matrix metalloproteinase-9 (MMP-9), MMP9, 92 kDa gelatinase, 92 kDa type IV collagenase, Gelatinase B (GELB), CLG4B, EC:3.4.24.35, UniProtKB# P14780

Principle of Method

The microtiter plate is coated with the antibody specifically binding the Matrix metalloproteinase 9. The human serum or plasma is incubated in the plate with the capture antibody.

The specimen is washed out and the specifically bound protein is incubated with biotin-labelled detection antibody. Following another washing step, StreptavidinHRP conjugate is added into the well. Unbound reagent is then washed out. Horseradish peroxidase (HRP) bound in the complex reacts with the chromogenic substrate (TMB) creating the blue colour. The reaction is stopped by addition of STOP solution (H2SO4).

The absorbance values are measured at 450 nm (optionally 450/630 nm) and are proportional to the concentration of MMP-9 in the specimen. The concentration of MMP-9 in unknown samples is determined from the calibration curve which is created by plotting the absorbance values against the standard concentration values.

Sample Types

Serum, Plasma

Typical Standard Curve

Assay Range: 0.625 ng/mL - 20 ng/mL

Sensitivity

The limit of detection, defined as a concentration of human MMP-9 giving absorbance higher than absorbance of blank +3 standard deviations, is better than 0.16 ng/mL of sample.

Precision

Intra-Assay

Test sample	Mean (ng/mL)	SD	CV
1	527	9.7	2%
2	224	6.7	3%

Inter-Assay (Run-to-Run)

Test sample	Mean (ng/mL)	SD	CV
1	389	31	8%
2	255	14	6%

Accuracy

Dilution Linearity:

Test sample	Dilution	Measured Concentration (ng/mL)	Expected Concentration (ng/mL)	Yield
1		519	-	-
1	2x	247	260	95%
1	4x	123	130	95%
1	8x	59	65	91%
2		582	-	-
2	2x	289	291	99%
2	4x	151	145	104%
2	8x	69	73	95%

Spiking Recovery:

Test sample	Spike (ng/mL)	Measured Concentration (ng/mL)	Expected Concentration (ng/mL)	Yield
1	-	223	-	-
1	250.0	480	473	102%
1	125.0	357	348	103%
1	62.5	290	285	102%

Storage

The kit must be stored at 2–8 °C. The opened components can be stored for one week at 2–8 °C.

Usage

This assay is for in vitro research use only and is not for use in diagnostic procedures.

Resources

Product Datasheet (PDS)

Summary References

Łukaszewicz-Zając M, Mroczko B, Słowik A. Matrix metalloproteinases (MMPs) and their tissue inhibitors (TIMPs) in amyotrophic lateral sclerosis (ALS). J Neural Transm (Vienna). 2014 Nov;121(11):1387-97. doi: 10.1007/s00702-014- 1205-3. Epub 2014 Jul 22. PMID: 25047909; PMCID: PMC4210652.
Nandi SS, Katsurada K, Sharma NM, Anderson DR, Mahata SK, Patel KP. MMP9 inhibition increases autophagic flux in chronic heart failure. Am J Physiol Heart Circ Physiol. 2020 Dec 1;319(6):H1414-H1437. doi: 10.1152/ajpheart.00032.2020. Epub 2020 Oct 16. PMID: 33064567; PMCID: PMC7792705.
Yadav SK, Kambis TN, Kar S, Park SY, Mishra PK. MMP9 mediates acute hyperglycemia-induced human cardiac stem cell death by upregulating apoptosis and pyroptosis in vitro. Cell Death Dis. 2020 Mar 13;11(3):186. doi: 10.1038/s41419-020- 2367-6. PMID: 32170070; PMCID: PMC7070071.
Snitker S, Xie K, Ryan KA, Yu D, Shuldiner AR, Mitchell BD, Gong DW. Correlation of circulating MMP-9 with white blood cell count in humans: effect of smoking. PLoS One. 2013 Jun 25;8(6):e66277. doi: 10.1371/journal.pone.0066277. PMID: 23825535; PMCID: PMC3692499.