Rat Total MMP-9 Quantikine ELISA Kit

R&D Systems | Catalog # RMP900

R&D Systems
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Key Product Details

Assay Length

4.5 hours

Sample Type & Volume Required Per Well

Cell Culture Supernates (50 µL), Serum (10 µL), Platelet-poor Heparin Plasma (10 µL)

Sensitivity

0.028 ng/mL

Assay Range

0.156-10 ng/mL
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Product Summary for Rat Total MMP-9 Quantikine ELISA Kit

The Quantikine Rat Total MMP-9 Immunoassay is a 4.5 hour solid phase ELISA designed to measure total rat MMP-9 (Pro-, active, and TIMP-complexed MMP-9) in cell culture supernates, serum, and plasma. It contains NS0-expressed recombinant rat MMP-9 and antibodies raised against the recombinant factor. This immunoassay has been shown to accurately quantitate the recombinant protein. Results obtained using natural rat MMP-9 showed dose-response curves that were parallel to the standard curves obtained using the Quantikine kit standards. These results indicate that this kit can be used to determine relative mass values for natural rat MMP-9.

Product Specifications

Assay Type

Solid Phase Sandwich ELISA

Format

96-well strip plate

Measurement

Quantitative ELISA

Detection Method

Colorimetric - 450nm (TMB)

Conjugate

HRP

Species

Rat

Specificity

Natural and recombinant rat MMP-9 (Pro-, active, and TIMP-complexed)

Cross-reactivity

< 0.5% cross-reactivity observed with available related molecules. < 50% cross-species reactivity observed with species tested.

Interference

No significant interference observed with available related molecules.

Precision

Intra-Assay Precision (Precision within an assay) Three samples of known concentration were tested on one plate to assess intra-assay precision.

Inter-Assay Precision (Precision between assays) Three samples of known concentration were tested in separate assays to assess inter-assay precision.

Cell Culture Supernates, Platelet-poor Heparin Plasma, Serum

Intra-Assay Precision Inter-Assay Precision
Sample 1 2 3 1 2 3
n 20 20 20 20 20 20
Mean (ng/mL) 0.39 1.22 5.27 0.51 1.18 3.55
Standard Deviation 0.027 0.068 0.238 0.035 0.046 0.153
CV% 6.9 5.6 4.5 6.9 3.9 4.3

Recovery for Rat Total MMP-9 Quantikine ELISA Kit

The recovery of rat MMP-9 spiked into various matrices was evaluated.

Sample Type Average % Recovery Range %
Cell Culture Samples (n=4) 94 87-107
Platelet-poor Heparin Plasma (n=4) 92 80-103
Serum (n=4) 97 90-115

Linearity

To assess the linearity of the assay, samples containing high concentrations of rat MMP-9 were serially diluted with Calibrator Diluent to produce samples with values within the dynamic range of the assay.

Rat Total MMP-9 ELISA Linearity

Scientific Data Images for Rat Total MMP-9 Quantikine ELISA Kit

Rat Total MMP-9 ELISA Standard Curve

Rat Total MMP-9 ELISA Standard Curve

Preparation and Storage

Shipping

The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below.

Stability & Storage

Store the unopened product at 2 - 8 °C. Do not use past expiration date.

Background: MMP-9

MMP-9 (Gelatinase B) is a metalloproteinase that degrades multiple extracellular matrix components including Gelatin, Collagens III, IV, V, and XI, Elastin, Nidogen-1, and Vitronectin. It also cleaves a variety of chemokines and growth factors (e.g. IL-1 beta, CXCL8/IL-8, CXCL7, CXCL4, CXCL1, Latent TGF-beta, membrane bound TNF-alpha, VEGF, and FGF basic), Amyloid beta peptide, Substance P, and Myelin Basic Protein. MMP-9 can trigger signaling through various transmembrane proteins or inhibit signaling by inducing their shedding from the cell surface (e.g. CD44, E-Cadherin, Integrins, ICAM-1, and CD25/IL-2 R alpha). The activity of MMP-9 is regulated by its association with the TIMP-1 protease inhibitor. Circulating levels of MMP-9 are elevated in many inflammatory disorders including intraluminal thrombus formation, atherosclerosis, Crohn’s disease, hepatitis C virus infection, colorectal cancer, and Duchenne muscular dystrophy.

Long Name

Matrix Metalloproteinase 9

Alternate Names

CLG4B, Gelatinase B, GELB, MANDP2, MMP9

Entrez Gene IDs

4318 (Human); 17395 (Mouse); 81687 (Rat); 102117693 (Cynomolgus Monkey)

Gene Symbol

MMP9

Additional MMP-9 Products

Product Documents for Rat Total MMP-9 Quantikine ELISA Kit

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot or batch number in the search box below.

Note: Certificate of Analysis not available for kit components.

Product Specific Notices for Rat Total MMP-9 Quantikine ELISA Kit

For research use only

⚠ WARNING: This product can expose you to chemicals including N,N-Dimethylforamide, which is known to the State of California to cause cancer. For more information, go to www.P65Warnings.ca.gov.

Citations for Rat Total MMP-9 Quantikine ELISA Kit

Customer Reviews for Rat Total MMP-9 Quantikine ELISA Kit (3)

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  • Rat Total MMP-9 Quantikine ELISA Kit
    Name: Anonymous
    Sample Tested: Serum and Rat serum
    Verified Customer | Posted 11/06/2020
    I followed the recommended 10-fold dilution and the kit worked perfectly fine.
  • Rat Total MMP-9 Quantikine ELISA Kit
    Name: Veena Rao
    Sample Tested: EDTA Plasma
    Verified Customer | Posted 06/09/2019
    Rat Total MMP-9 Quantikine ELISA Kit RMP900
  • Rat Total MMP-9 Quantikine ELISA Kit
    Name: BENARD OGOLA
    Sample Tested: Human cell conditioned medium
    Verified Customer | Posted 05/11/2017
    Easy to follow and extrapolate graph for experiments

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Protocols

View specific protocols for Rat Total MMP-9 Quantikine ELISA Kit (RMP900):

Refer to the product for complete assay procedure.

Bring all reagents and samples to room temperature before use. It is recommended that all samples, standards, and controls be assayed in duplicate.
  1.   Prepare all reagents, standard dilutions, and samples as directed in the product insert.
  2.   Remove excess microplate strips from the plate frame, return them to the foil pouch containing the desiccant pack, and reseal.
    3. 50 µL Assay Diluent
  3.   Add 50 µL of Assay Diluent to each well.
    4. 50 µL Standard, Control, or Sample
  4.   Add 50 µL of Standard, control, or sample to each well. Cover with a plate sealer, and incubate at room temperature for 2 hours.
  5.   Aspirate each well and wash, repeating the process 4 times for a total of 5 washes.
    6. 100 µL Conjugate
  6.   Add 100 µL of Conjugate to each well. Cover with a new plate sealer, and incubate at room temperature for 2 hours.
  7.   Aspirate and wash 5 times.
    8. 100 µL Substrate Solution
  8.   Add 100 µL Substrate Solution to each well. Incubate at room temperature for 30 minutes. PROTECT FROM LIGHT.
    9. 100 µL Stop Solution
  9.   Add 100 µL of Stop Solution to each well. Read at 450 nm within 30 minutes. Set wavelength correction to 540 nm or 570 nm.

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