TIMP1 Kit ELISA

N° du produit ABIN625179

Détail du produit TIMP1 Kit ELISA

Antigène: TIMP1 (TIMP Metallopeptidase Inhibitor 1 (TIMP1))

Reactivité: Souris

Méthode de détection: Colorimetric

Type de méthode: Sandwich ELISA

Gamme de detection: 3-1000 pg/mL

Seuil minimal de détection: 3 pg/mL

Application: ELISA

Fonction: Mouse TIMP-1 ELISA Kit for cell culture supernatants, plasma, and serum samples.

Type d'échantillon: Plasma, Cell Culture Supernatant, Serum

Analytical Method: Quantitative

Specificité: This ELISA kit shows no cross-reactivity with any of the 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-6, 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, MCSF, MIG, MIP-1 alpha, MIP-1 gamma, MIP-2, MIP-3 beta, MIP-3 alpha, PF-4, P-Selectin, RANTES, SCF, SDF-1 alpha, TARC, TCA-3, TECK, TNF-alpha, TNF RI, TNF RII, TPO, VCAM-1, VEGF.

Sensibilité: < 3 pg/mL

Attributs du produit:

• 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

Ingrédients:

• 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

Matériel non inclus:

• 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

Information d'application

Indications d'application: Recommended Dilution for serum and plasma samples10 - 100 fold

Volume d'échantillon: 100 μL

Plaque: Pre-coated

Protocole:

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.

Préparation des réactifs:

1. Bring all reagents and samples to room temperature (18 - 25 °C) before use.
   2. Sample dilution: If your samples need to be diluted, Assay Diluent A (Item D) should be used for dilution of serum/plasma samples. 1x Assay Diluent B (Item E) should be used for dilution of culture supernatants. Suggested dilution for normal serum/plasma: 10-100 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 B should be diluted 5-fold with deionized or distilled water.
   4. Preparation of standard: Briefly spin the vial of Item C. Add 400 µL Assay Diluent A (for serum/plasma samples) or 1x Assay Diluent B (for cell culture medium) into Item C vial to prepare a 50 ng/mL standard. Dissolve the powder thoroughly by a gentle mix. Add 20 µL TIMP-1 standard from the vial of Item C, into a tube with 980.0 µL Assay Diluent A or 1x Assay Diluent B to prepare a 1,000 pg/mL stock standard solution. Pipette 400 µL Assay Diluent A or 1x Assay Diluent B into each tube. Use the stock standard solution to produce a dilution series . Mix each tube thoroughly before the next transfer. Assay Diluent A or 1x Assay Diluent B serves as the zero standard (0 pg/mL). 200 µL 200 µL 200 µL 200 µL 20 µL standard +980.0 µL 200myl 200 µL 1000 333.3 111.1 37.04 12.35 4.12 1.37 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 B 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 B 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 120-fold with 1x Assay Diluent B. For example: Briefly spin the vial (Item G) and pipette up and down to mix gently . Add 100 µL of HRP-Streptavidin concentrate into a tube with 12 ml 1x Assay Diluent B to prepare a final 120 fold diluted HRP-Streptavidin solution. Mix well.

Procédure de l'essai:

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.

Calcul des résultats:

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 A Mouse TIMP-1 concentration (pg/mL) 0.1 1 10 100 1000 10000 O D =4 50 n m 0.01 0.1 1 10 Assay Diluent B Mouse TIMP-1 concentration (pg/mL) 0.1 1 10 100 1000 10000 O D =4 50 n m 0.001 0.01 0.1 1 10
Sensitivity: The minimum detectable dose of TIMP-1 is typically less than 3 pg/mL.
Recovery: Recovery was determined by spiking various levels of mouse TIMP-1 into mouse serum, plasma and cell culture media. Mean recoveries are as follows: Sample Type Average % Recovery Range ( %) Serum 94.95 80-101 Plasma 92.28 78-100 Cell culture media 102.53 89-111
Linearity: Sample Type Serum Plasma Cell Culture Media 1:2 Average % of Expected 83 87 94 Range ( %) 78-101 80-102 84-105 1:4 Average % of Expected 89 91 102 Range ( %) 80-104 83-104 90-109
Reproducibility: Intra-Assay: CV<10 % Inter-Assay: CV<12 %

Précision du teste: Intra-Assay: CV< 10 % Inter-Assay: CV< 12 %

Restrictions: For Research Use only

Stockage

Conseil sur la manipulation: Avoid repeated freeze-thaw cycles.

Stock: -20 °C

Stockage commentaire: 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.

Date de péremption: 6 months

Références pour TIMP1 Kit ELISA (ABIN625179)

Carroll-Portillo, Cannon, te Riet, Holmes, Kawakami, Kawakami, Cambi, Lidke: "Mast cells and dendritic cells form synapses that facilitate antigen transfer for T cell activation." dans: The Journal of cell biology, Vol. 210, Issue 5, pp. 851-64, (2015) (PubMed).

Song, Hu, He, Wang, Liu, Ma, Lin, Chen: "Novel biomarkers for early prediction of sepsis-induced disseminated intravascular coagulation in a mouse cecal ligation and puncture model." dans: Journal of inflammation (London, England), Vol. 10, Issue 1, pp. 7, (2013) (PubMed).

Raghu, Nalla, Gondi, Gujrati, Dinh, Rao: "uPA and uPAR shRNA inhibit angiogenesis via enhanced secretion of SVEGFR1 independent of GM-CSF but dependent on TIMP-1 in endothelial and glioblastoma cells." dans: Molecular oncology, Vol. 6, Issue 1, pp. 33-47, (2012) (PubMed).

Spruss, Kanuri, Stahl, Bischoff, Bergheim: "Metformin protects against the development of fructose-induced steatosis in mice: role of the intestinal barrier function." dans: Laboratory investigation; a journal of technical methods and pathology, Vol. 92, Issue 7, pp. 1020-32, (2012) (PubMed).

Kisarova, Korolenko: "Tissue inhibitors of matrix metalloproteinases 1 and 2 and matrix metalloproteinase activity in the serum and lungs of mice with lewis lung carcinoma." dans: Bulletin of experimental biology and medicine, Vol. 153, Issue 6, pp. 874-7, (2012) (PubMed).

Guzowska-Nowowiejska, Fiedorowicz, Plader: "Cucumber, melon, pumpkin, and squash: are rules of editing in flowering plants chloroplast genes so well known indeed?" dans: Gene, Vol. 434, Issue 1-2, pp. 1-8, (2009) (PubMed).

Détail du antigène

Antigène: TIMP1 (TIMP Metallopeptidase Inhibitor 1 (TIMP1))

Autre désignation: TIMP-1 (TIMP1 Produits)

Synonymes: TIMP-1 Kit ELISA, TIMP1 Kit ELISA, DKFZp468A0912 Kit ELISA, CLGI Kit ELISA, EPA Kit ELISA, EPO Kit ELISA, HCI Kit ELISA, TIMP Kit ELISA, Timp Kit ELISA, Clgi Kit ELISA, TIMP metallopeptidase inhibitor 1 Kit ELISA, tissue inhibitor of metalloproteinase 1 Kit ELISA, TIMP1 Kit ELISA, Timp1 Kit ELISA

Sujet: Metalloproteinase inhibitor 1 (Collagenase inhibitor 16C8 fibroblast) (Erythroid-potentiating activity) (EPA) (TPA-S1) (TPA-induced protein) (Tissue inhibitor of metalloproteinases 1) (TIMP-1)

ID gène: 21857

UniProt: P12032