MMP2 ELISA Kit

Produktnummer ABIN625167

Produktdetails MMP2 ELISA Kit

Target: MMP2 (Matrix Metalloproteinase 2 (MMP2))

Reaktivität: Maus

Nachweismethode: Colorimetric

Methodentyp: Sandwich ELISA

Detektionsbereich: 25-20000 pg/mL

Untere Nachweisgrenze: 25 pg/mL

Applikation: ELISA

Verwendungszweck: Mouse MMP-2 ELISA Kit for cell culture supernatants, plasma, and serum samples.

Proben: Plasma, Cell Culture Supernatant, Serum

Analytische Methode: Quantitative

Spezifität: This ELISA kit shows no cross-reactivity with the following cytokines tested: Mouse CD30, L CD30, T CD40, CRG-2, CTACK, CXCL16, Eotaxin , Eotaxin-2, Fas Ligand, Fractalkine, GCSF, GM-CFS, IFN- gamma, IGFBP-3, IGFBP-5, IGFBP-6, IL-1 alpha, IL-1 beta, IL-2, IL-3, IL-3 Rb, IL-4, IL-5, IL-9, IL-10, IL-12 p40/p70, IL-12 p70, IL-13, IL-17, KC, Leptin R, LEPTIN(OB), LIX, L-Selectin, Lymphotactin, MCP-1, MCP- 5, M-CSF, MIG, MIP-1 alpha, MIP-1 gamma, MIP-2, MIP-3 beta, MIP-3 alpha, MMP-3, proMMP-9, PF-4, P-Selectin, RANTES, SCF, SDF-1 alpha, TARC, TCA-3, TECK, TIMP-1, TNF-alpha, TNF RI, TNF RII, TPO, VCAM-1, VEGF.

Sensitivität: < 25 pg/mL

Produktmerkmale:

• Strip plates and additional reagents allow for use in multiple experiments
• Quantitative protein detection
• Establishes normal range
• The best products for confirmation of antibody array data

Bestandteile:

• Pre-Coated 96-well Strip Microplate
• Wash Buffer
• Stop Solution
• Assay Diluent(s)
• Lyophilized Standard
• Biotinylated Detection Antibody
• Streptavidin-Conjugated HRP
• TMB One-Step Substrate

Benötigtes Material:

• Distilled or deionized water
• Precision pipettes to deliver 2 μL to 1 μL volumes
• Adjustable 1-25 μL pipettes for reagent preparation
• 100 μL and 1 liter graduated cylinders
• Tubes to prepare standard and sample dilutions
• Absorbent paper
• Microplate reader capable of measuring absorbance at 450nm
• Log-log graph paper or computer and software for ELISA data analysis

Anwendungsinformationen

Applikationshinweise: Recommended Dilution for serum and plasma samples100 - 1,000 fold

Probenmenge: 100 μL

Plattentyp: Pre-coated

Protokoll:

1. Prepare all reagents, samples and standards as instructed in the manual.
2. Add 100 μL of standard or sample to each well.
3. Incubate 2.5 h at RT or O/N at 4 °C.
4. Add 100 μL of prepared biotin antibody to each well.
5. Incubate 1 h at RT.
6. Add 100 μL of prepared Streptavidin solution to each well.
7. Incubate 45 min at RT.
8. Add 100 μL of TMB One-Step Substrate Reagent to each well.
9. Incubate 30 min at RT.
10. Add 50 μL of Stop Solution to each well.
11. Read at 450 nm immediately.

Aufbereitung der Reagenzien:

1. Bring all reagents and samples to room temperature (18 - 25 °C) before use.
   2. Sample dilution: If your samples need to be diluted, 1x Assay Diluent (Item E) should be used for dilution of serum/plasma/culture supernatants. Suggested dilution for normal serum/plasma: 100-1000 fold. Please note that levels of the target protein may vary between different specimens. Optimal dilution factors for each sample must be determined by the investigator.
   3. Assay Diluent (Item E) should be diluted 5-fold with deionized or distilled water before use.
   4. Preparation of standard: Briefly spin the vial of Item C. Add 400 µL 1x Assay Diluent (Item E) into Item C vial to prepare a 100 ng/mL standard solution. Dissolve the powder thoroughly by a gentle mix. Add 100 µL MMP-2 standard from the vial of tem C, into a tube with 400 µL 1x Assay Diluent to prepare a 20,000 pg/mL standard solution. Pipette 400myl 1x Assay Diluent into each tube. Use the 20,000 pg/mL standard solution to produce a dilution series . Mix each tube thoroughly before the next transfer. 1x Assay Diluent serves as the zero standard (0 pg/mL). 200 µL 200 µL 200 µL 200 µL 2200 µL 100 µL standard + 400 µL 200myl 20,000 6667 2222 740.7 246.9 82.30 27.43 0 pg/mL pg/mL pg/mL pg/mL pg/mL pg/mL pg/mL pg/mL
   5. If the Wash Concentrate (20x) (Item B) contains visible crystals, warm to room temperature and mix gently until dissolved. Dilute 20 ml of Wash Buffer Concentrate into deionized or distilled water to yield 400 ml of 1x Wash Buffer.
   6. Briefly spin the Detection Antibody vial (Item F) before use. Add 100 µL of 1x Assay Diluent into the vial to prepare a detection antibody concentrate. Pipette up and down to mix gently (the concentrate can be stored at 4 °C for 5 days). The detection antibody concentrate should be diluted 80-fold with 1x Assay Diluent and used in step 4 of Part VI Assay Procedure.
   7. Briefly spin the HRP-Streptavidin concentrate vial (Item G) and pipette up and down to mix gently before use. HRP-Streptavidin concentrate should be diluted 400-fold with 1x Assay Diluent. For example: Briefly spin the vial (Item G) and pipette up and down to mix gently . Add 30 µL of HRP-Streptavidin concentrate into a tube with 12 ml 1x Assay Diluent to prepare a 400-fold diluted HRP-Streptavidin solution (don't store the diluted solution for next day use). Mix well.

Testdurchführung:

1. Bring all reagents and samples to room temperature (18 - 25 °C) before use. It is recommended that all standards and samples be run at least in duplicate.
   2. Add 100 µL of each standard (see Reagent Preparation step 2) and sample into appropriate wells. Cover well and incubate for 2.5 hours at room temperature or over night at 4 °C with gentle shaking.
   3. Discard the solution and wash 4 times with 1x Wash Solution. Wash by filling each well with Wash Buffer (300 myl) using a multi-channel Pipette or autowasher. Complete removal of liquid at each step is essential to good performance. After the last wash, remove any remaining Wash Buffer by aspirating or decanting. Invert the plate and blot it against clean paper towels.
   4. Add 100 µL of 1x prepared biotinylated antibody (Reagent Preparation step 6) to each well. Incubate for 1 hour at room temperature with gentle shaking.
   5. Discard the solution. Repeat the wash as in step
   6. Add 100 µL of prepared Streptavidin solution (see Reagent Preparation step 7) to each well. Incubate for 45 minutes at room temperature with gentle shaking.
   7. Discard the solution. Repeat the wash as in step
   8. Add 100 µL of TMB One-Step Substrate Reagent (Item H) to each well. Incubate for 30 minutes at room temperature in the dark with gentle shaking.
   9. Add 50 µL of Stop Solution (Item I) to each well. Read at 450 nm immediately.

Ergebnisberechnung:

Calculate the mean absorbance for each set of duplicate standards, controls and samples, and subtract the average zero standard optical density. Plot the standard curve on log-log graph paper or using Sigma plot software, with standard concentration on the x-axis and absorbance on the y-axis. Draw the best-fit straight line through the standard points.
Typical Data: These standard curves are for demonstration only. A standard curve must be run with each assay. Assay Diluent Mouse MMP-2 concentration (pg/mL) O D =4 50 n m 0.01 0.1 1 10 10 100 1,000 10,000 100,000
Sensitivity: The minimum detectable dose of MMP-2 is typically less than 25 pg/mL.
Recovery: Recovery was determined by spiking various levels of MMP-2 into normal mouse serum, plasma and cell culture media. Mean recoveries are as follows: Sample Type Average % Recovery Range ( %) Serum 107.6 96-127 Plasma 105.6 95-120 Cell culture media 94.21 86-104
Linearity: Sample Type Serum Plasma Cell Culture Media 1:2 Average % of Expected 97.09 102.5 103.3 Range ( %) 89-107 93-111 94-110 1:4 Average % of Expected 74.61 84.44 75.83 Range ( %) 68-86 76-97 69-89
Reproducibility: Intra-Assay: CV<10 % Inter-Assay: CV<12 %

Testpräzision: Intra-Assay: CV< 10 % Inter-Assay: CV< 12 %

Beschränkungen: Nur für Forschungszwecke einsetzbar

Handhabung

Handhabung: Avoid repeated freeze-thaw cycles.

Lagerung: -20 °C

Informationen zur Lagerung: The entire kit may be stored at -20°C for up to 1 year from the date of shipment. Avoid repeated freeze-thaw cycles. The kit may be stored at 4°C for up to 6 months. For extended storage, it is recommended to store at -80°C.

Haltbarkeit: 6 months

Referenzen für MMP2 ELISA Kit (ABIN625167)

Sun, Yu, Yu, Honglin, Ni, Zhang, Guo, He, Xue, Ni, Li, Feng, Chen, Shao, Chai, Li: "Inhibition of the Activation and Recruitment of Microglia-Like Cells Protects Against Neomycin-Induced Ototoxicity." in: Molecular neurobiology, (2014) (PubMed).

Thiyagarajan, Fismen, Seredkina, Jacobsen, Elung-Jensen, Kamper, Fenton, Rekvig, Mortensen: "Silencing of renal DNaseI in murine lupus nephritis imposes exposure of large chromatin fragments and activation of Toll like receptors and the Clec4e." in: PLoS ONE, Vol. 7, Issue 3, pp. e34080, (2012) (PubMed).

Xu, Ehrman, Graham, Eagleton: "Interleukin-5 is a potential mediator of angiotensin II-induced aneurysm formation in apolipoprotein E knockout mice." in: The Journal of surgical research, Vol. 178, Issue 1, pp. 512-8, (2012) (PubMed).

Yang, Wang, Zhou, Wu, Xing: "TNF-α stimulates MMP-2 and MMP-9 activities in human corneal epithelial cells via the activation of FAK/ERK signaling." in: Ophthalmic research, Vol. 48, Issue 4, pp. 165-70, (2012) (PubMed).

Chen, Lu, Gao, Wu, Wang, Shi: "Anti-angiogenesis effect of essential oil from Curcuma zedoaria in vitro and in vivo." in: Journal of ethnopharmacology, Vol. 133, Issue 1, pp. 220-6, (2010) (PubMed).

Details zu MMP2

Target: MMP2 (Matrix Metalloproteinase 2 (MMP2))

Andere Bezeichnung: MMP-2 (MMP2 Produkte)

Synonyme: 2-MMP ELISA Kit, CG1794 ELISA Kit, DM2-MMP ELISA Kit, Dm2-MMP ELISA Kit, Dmel\\CG1794 ELISA Kit, MMP2 ELISA Kit, anon-WO0118547.84 ELISA Kit, dm-2MMP ELISA Kit, dmmp2 ELISA Kit, l(2)02353 ELISA Kit, mmp2 ELISA Kit, wu:fa99h12 ELISA Kit, wu:fk89d01 ELISA Kit, fgmmp-2 ELISA Kit, Mmp-2 ELISA Kit, LOC100135793 ELISA Kit, Clg4a ELISA Kit, GelA ELISA Kit, MMP-2 ELISA Kit, CLG4 ELISA Kit, CLG4A ELISA Kit, MMP-II ELISA Kit, MONA ELISA Kit, TBE-1 ELISA Kit, matrix metallopeptidase 2 ELISA Kit, Matrix metalloproteinase 2 ELISA Kit, matrix metalloproteinase 2 ELISA Kit, matrix metalloproteinase-2 ELISA Kit, MMP2 ELISA Kit, Mmp2 ELISA Kit, mmp2 ELISA Kit, LOC657982 ELISA Kit, LOC100135793 ELISA Kit

Hintergrund: 72 kDa type IV collagenase (EC 3.4.24.24) (72 kDa gelatinase) (Gelatinase A) (Matrix metalloproteinase-2) (MMP-2)

Gen-ID: 17390

UniProt: P33434

Pathways: Activation of Innate immune Response