Chemokine (C-C Motif) Ligand 2 (CCL2) ELISA Kit

Details zu Produkt Nr. ABIN625210, Anbieter: Anmelden zum Anzeigen
Antigen
  • MCP-1
  • Scya2
  • Sigje
  • GDCF-2
  • HC11
  • HSMCR30
  • MCAF
  • MCP1
  • SCYA2
  • SMC-CF
  • AI323594
  • JE
  • MCP-1A
  • MCP1A
  • chemokine (C-C motif) ligand 2
  • macrophage cationic peptide 1
  • mast cell proteinase-1
  • CCL2
  • Ccl2
  • MCP-1
  • MCP1
Alternativen
Ratte (Rattus) Chemokine (C-C Motif) Ligand 2 ELISA Kit
Reaktivität
Ratte (Rattus)
Alternativen
Kits mit alternativen Reaktivitäten:
52
36
28
17
15
14
9
8
8
3
2
2
2
1
1
Methodentyp
Sandwich ELISA
Detektionsbereich
15-18000 pg/mL
Untere Nachweisgrenze
15 pg/mL
Applikation
ELISA
Optionen
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Verwendungszweck Rat MCP-1 (CCL2) ELISA Kit for cell culture supernatants, plasma, and serum samples.
Proben Plasma, Cell Culture Supernatant, Serum
Analytische Methode Quantitative
Nachweismethode Colorimetric
Spezifität This ELISA kit shows no cross-reactivity with any of the cytokines tested: rat CINC-2, CINC-3, CNTF, IL-1 alpha, IL-1 beta, IL-4, IL-6, IL-10, GM-CSF, IFN-gamma, Leptin, Lix, MIP-3 alpha, beta-NGF, TIMP-1, TNF-alpha, VEGF.
Sensitivität < 15 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
Andere Bezeichnung MCP-1 / CCL2 (CCL2 ELISA Kit Abstract)
Hintergrund C-C motif chemokine 2 (Immediate-early serum-responsive protein JE) (Monocyte chemoattractant protein 1) (Monocyte chemotactic protein 1) (MCP-1) (Small-inducible cytokine A2)
Gen-ID 24770
UniProt P14844
Forschungsgebiet Chemokines
Pathways Cellular Response to Molecule of Bacterial Origin, Positive Regulation of Immune Effector Process, ER-Nucleus Signaling
Applikationshinweise Recommended Dilution for serum and plasma samples500 - 10,000 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, 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: 500-10,000 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 200 µL MCP-1 standard from the vial of Item C, into a tube with 355.6 µL Assay Diluent A or 1x Assay Diluent B to prepare a 18,000 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 200myl 200 µL 200 µL 200 µL 200 µL 200 µL standard + 355.6 µL 18,000 6,000 2,000 666.7 222.2 74.07 24.67 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 65-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 100-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 10 ml 1x Assay Diluent B to prepare a 100 fold diluted HRP-Streptavidin solution. 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 A Rat MCP-1 concentration (pg/mL) O D =4 50 (n m ) 0.01 0.1 1 10 10 100 1,000 10,000 100,000 Assay Diluent B Rat MCP-1 concentration (pg/mL) O D =4 50 (n m ) 0.001 0.01 0.1 1 10 10 100 1,000 10,000 100,000
Sensitivity: The minimum detectable dose of MCP-1 is typically less than 15 pg/mL.
Recovery: Recovery was determined by spiking various levels of rat MCP-1 into rat serum, plasma and cell culture media. Mean recoveries are as follows: Sample Type Average % Recovery Range ( %) Serum 95.39 84-103 Plasma 92.44 83-103 Cell culture media 101.4 86-105
Linearity: Sample Type Serum Plasma Cell Culture Media 1:2 Average % of Expected 95 92 97 Range ( %) 84-103 82-102 85-104 1:4 Average % of Expected 97 97 101 Range ( %) 85-105 84-104 86-107
Reproducibility: Intra-Assay: CV<10 % Inter-Assay: CV<12 %

Testpräzision Intra-Assay: CV< 10 % Inter-Assay: CV< 12 %
Beschränkungen Nur für Forschungszwecke einsetzbar
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
Bilder des Herstellers
ELISA image for Chemokine (C-C Motif) Ligand 2 (CCL2) ELISA Kit (ABIN625210) Chemokine (C-C Motif) Ligand 2 (CCL2) ELISA Kit
Produkt verwendet in: Alique, Sánchez-López, Rayego-Mateos, Egido, Ortiz, Ruiz-Ortega: "Angiotensin II, via angiotensin receptor type 1/nuclear factor-?B activation, causes a synergistic effect on interleukin-1-?-induced inflammatory responses in cultured mesangial cells." in: Journal of the renin-angiotensin-aldosterone system : JRAAS, Vol. 16, Issue 1, pp. 23-32, 2015 (PubMed).

Zhang, Zhang, Tsui: "Mesenteric lymph duct drainage attenuates acute lung injury in rats with severe intraperitoneal infection." in: Inflammation, Vol. 38, Issue 3, pp. 1239-49, 2015 (PubMed).

Yan, Fang, Yang, Li, Lu, Cheng: "Anti-diabetic nephropathy compounds from Cinnamomum cassia." in: Journal of ethnopharmacology, Vol. 165, pp. 141-7, 2015 (PubMed).

Kaur, Patro, Tikoo, Sandhir: "Curcumin attenuates inflammatory response and cognitive deficits in experimental model of chronic epilepsy." in: Neurochemistry international, Vol. 89, pp. 40-50, 2015 (PubMed).

Ikutomi, Sahara, Nakajima, Minami, Morita, Hirata, Komuro, Nakamura, Sata: "Diverse contribution of bone marrow-derived late-outgrowth endothelial progenitor cells to vascular repair under pulmonary arterial hypertension and arterial neointimal formation." in: Journal of molecular and cellular cardiology, Vol. 86, pp. 121-35, 2015 (PubMed).

Wu, Lv, Chen, Ma, Shao, Wang: "The function of miR-199a-5p/Klotho regulating TLR4/NF-?B p65/NGAL pathways in rat mesangial cells cultured with high glucose and the mechanism." in: Molecular and cellular endocrinology, Vol. 417, pp. 84-93, 2015 (PubMed).

Lebrun-Harris, Fiore, Tomoyasu, Ngo-Metzger: "Cigarette Smoking, Desire to Quit, and Tobacco-Related Counseling Among Patients at Adult Health Centers." in: American journal of public health, 2014

Lebrun-Harris, Fiore, Tomoyasu, Ngo-Metzger: "Cigarette Smoking, Desire to Quit, and Tobacco-Related Counseling Among Patients at Adult Health Centers." in: American journal of public health, 2014

Lebrun-Harris, Fiore, Tomoyasu, Ngo-Metzger: "Cigarette Smoking, Desire to Quit, and Tobacco-Related Counseling Among Patients at Adult Health Centers." in: American journal of public health, 2014

Yang, Sun, Bhaumik, Li, Baumann, Lin, Zhang, Lin, Dunn, Liu, Shie, Lee, Wills-Karp, Chougnet, Kallapur, Lewkowich, Lindquist, Murali-Krishna, Kuan: "Blocking lymphocyte trafficking with FTY720 prevents inflammation-sensitized hypoxic-ischemic brain injury in newborns." in: The Journal of neuroscience : the official journal of the Society for Neuroscience, Vol. 34, Issue 49, pp. 16467-81, 2014 (PubMed).

Smits, Ballotta, Driessen-Mol, Bouten, Baaijens: "Shear flow affects selective monocyte recruitment into MCP-1-loaded scaffolds." in: Journal of cellular and molecular medicine, Vol. 18, Issue 11, pp. 2176-88, 2014 (PubMed).

Hsu, Wang, Yang, Yang, Yang: "Effects of fenofibrate on adiponectin expression in retinas of streptozotocin-induced diabetic rats." in: Journal of diabetes research, Vol. 2014, pp. 540326, 2014 (PubMed).

Ergenoğlu, Yeniel, Erbaş, Aktuğ, Yildirim, Ulukuş, Taskiran: "Regression of endometrial implants by resveratrol in an experimentally induced endometriosis model in rats." in: Reproductive sciences (Thousand Oaks, Calif.), Vol. 20, Issue 10, pp. 1230-6, 2013

Ergenoğlu, Yeniel, Erbaş, Aktuğ, Yildirim, Ulukuş, Taskiran: "Regression of endometrial implants by resveratrol in an experimentally induced endometriosis model in rats." in: Reproductive sciences (Thousand Oaks, Calif.), Vol. 20, Issue 10, pp. 1230-6, 2013

Ergenoğlu, Yeniel, Erbaş, Aktuğ, Yildirim, Ulukuş, Taskiran: "Regression of endometrial implants by resveratrol in an experimentally induced endometriosis model in rats." in: Reproductive sciences (Thousand Oaks, Calif.), Vol. 20, Issue 10, pp. 1230-6, 2013

Ergeno?lu, Yeniel, Erba?, Aktu?, Yildirim, Uluku?, Taskiran: "Regression of endometrial implants by resveratrol in an experimentally induced endometriosis model in rats." in: Reproductive sciences (Thousand Oaks, Calif.), Vol. 20, Issue 10, pp. 1230-6, 2013 (PubMed).

Savary-Auzeloux, Magne, Migné, Oberli, Breuillé, Faure, Vidal, Perrot, Rémond, Combaret, Dardevet: "A dietary supplementation with leucine and antioxidants is capable to accelerate muscle mass recovery after immobilization in adult rats." in: PLoS ONE, Vol. 8, Issue 11, pp. e81495, 2013 (PubMed).

Schaal, Garg, Ghosh, Lovera, Lopez, Patel, Louro, Patel, Tuesta, Chan, Pearse: "The therapeutic profile of rolipram, PDE target and mechanism of action as a neuroprotectant following spinal cord injury." in: PLoS ONE, Vol. 7, Issue 9, pp. e43634, 2012 (PubMed).

Yamaleyeva, Guimaraes-Souza, Krane, Agcaoili, Gyabaah, Atala, Aboushwareb, Yoo: "Cell therapy with human renal cell cultures containing erythropoietin-positive cells improves chronic kidney injury." in: Stem cells translational medicine, Vol. 1, Issue 5, pp. 373-83, 2012 (PubMed).

Allgemeine Veröffentlichungen Lebrun-Harris, Fiore, Tomoyasu, Ngo-Metzger: "Cigarette Smoking, Desire to Quit, and Tobacco-Related Counseling Among Patients at Adult Health Centers." in: American journal of public health, 2014 (PubMed).