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

Details zu Produkt Nr. ABIN625159, 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
Maus Chemokine (C-C Motif) Ligand 2 ELISA Kit
Reaktivität
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35
29
17
15
14
9
8
8
3
2
2
2
1
1
Methodentyp
Sandwich ELISA
Detektionsbereich
3-2000 pg/mL
Untere Nachweisgrenze
3 pg/mL
Applikation
ELISA
Optionen
Hersteller
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Verwendungszweck Mouse MCP-1 (CCL2) ELISA Kit for cell culture supernatants, plasma, and serum samples.
Proben Serum, Plasma, Cell Culture Supernatant
Analytische Methode Quantitative
Nachweismethode Colorimetric
Spezifität This ELISA kit shows no cross-reactivity with any of the cytokines tested: Mouse 6Ckine, CTACK, Eotaxin, GCSF, GM-CSF, IL-2, IL-3, IL-4, IL-5, IL-6, IL-9, IL-10, IL-12p40, IL-12p70, IL-13, IL-17, IFN-gamma, KC, Leptin, MCP-5, MIP-1 alpha, MIP-2, MIP-3 beta, RANTES, SCF, sTNFri, TARC, TIMP-1, TNF-alpha, TPO, VEGF.
Sensitivität < 3 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 Monocyte chemoattractant protein-1 (MCP-1), also called monocyte chemotactic protein-1, belongs to the family of Chemokines. MCP-1 is expressed by monocytes, vascular endothelial cells, smooth muscle cells, glomerular mesangial cell, and osteoblastic cells. It has been shown to exhibit biological activities other than Chemotaxis. The activity of mouse MCP-1 has been shown to be mediated by the mouse CC chemokine receptor CCR2. The Mouse MCP-1 ELISA (Enzyme-Linked Immunosorbent Assay) kit is an in vitro enzyme-linked immunosorbent assay for the quantitative measurement of mouse MCP-1 in serum, plasma and cell culture supernatants. This assay employs an antibody specific for mouse MCP-1 coated on a 96-well plate. Standards and samples are pipetted into the wells and MCP-1 present in a sample is bound to the wells by the immobilized antibody. The wells are washed and biotinylated anti-mouse MCP-1 antibody is added. After washing away unbound biotinylated antibody, HRP-conjugated streptavidin is pipetted to the wells. The wells are again washed, a TMB substrate solution is added to the wells and color develops in proportion to the amount of MCP-1 bound. The Stop Solution changes the color from blue to yellow, and the intensity of the color is measured at 450 nm. Reproducibility: Intra-Assay: CV<10% Inter-Assay: CV<12%.
Gen-ID 20296
UniProt P10148
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 samples2 - 10 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: 2-10 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 and then 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 40 µL MCP-1 standard from the vial of Item C, into a tube with 960 µL Assay Diluent A or 1x Assay Diluent B to prepare a 2,000 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 200 µL 40 µL standard + 960 µL 200myl 2,000 666.7 222.2 74.07 24.69 8.23 2.74 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 100-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) before use. HRP-Streptavidin concentrate should be diluted 400-fold with 1x Assay Diluent B. 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 B 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 A Mouse MCP-1concentration (pg/mL) 1 10 100 1000 10000 O D =4 50 (n m ) 0.01 0.1 1 10 Assay Diluent B Mouse MCP-1concentration (pg/mL) 1 10 100 1000 10000 O D =4 50 (n m ) 0.01 0.1 1 10
Sensitivity: The minimum detectable dose of MCP-1 is typically less than 3 pg/mL.
Recovery: Recovery was determined by spiking various levels of mouse MCP-1 into mouse serum, plasma and cell culture media. Mean recoveries are as follows: Sample Type Average % Recovery Range ( %) Serum 94.28 83-103 Plasma 92.79 82-102 Cell culture media 95.29 84-104
Linearity: Sample Type Serum Plasma Cell Culture Media 1:2 Average % of Expected 93 92 94 Range ( %) 83-102 82-102 83-103 1:4 Average % of Expected 94 95 93 Range ( %) 83-102 84-103 82-103
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 (ABIN625159) Chemokine (C-C Motif) Ligand 2 (CCL2) ELISA Kit
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