CCL2 ELISA Kit

Produktnummer ABIN1112743

Maus CCL2 ELISA Kit, Colorimetric Assay für die Quantifizierung von Maus CCL2.

Kurzübersicht für CCL2 ELISA Kit (ABIN1112743)

Target: CCL2 (Chemokine (C-C Motif) Ligand 2 (CCL2))

Reaktivität: Maus

Nachweismethode: Colorimetric

Methodentyp: Sandwich ELISA

Detektionsbereich: 15.6 pg/mL - 1000 pg/mL

Applikation: ELISA

Proben: Cell Culture Supernatant, Plasma, Serum, Tissue Lysate

Produktdetails CCL2 ELISA Kit

Untere Nachweisgrenze: 15.6 pg/mL

Verwendungszweck: For quantitative detection of MCP-1 in mouse serum, plasma, tissue lysatesor cell culture supernatants.

Analytische Methode: Quantitative

Sensitivität: < 3 pg/mL

Bestandteile: 1. One 96-well plate pre-coated with anti-Mouse MCP-1 antibody 2. Lyophilized Mouse MCP-1 standards: 2 tubes (10ng / tube) 3. Sample / Standard diluent buffer: 30ml 4. Biotin conjugated anti-Mouse MCP-1 antibody (Concentrated): 130 µl.

Benötigtes Material:

1. 37 °C incubator
2. Microplate reader (wavelength: 450 nm)
3. Precise pipette and disposable pipette tips
4. Automated plate washer
5. ELISA shaker
6. 1.5 mL of Eppendorf tubes
7. Plate cover
8. Absorbent filter papers
9. Plastic or glass container with volume of above 1 L

Anwendungsinformationen

Applikationshinweise: Optimal working dilution should be determined by the investigator.

Kommentare:

This kit was based on sandwich enzyme-linked immune-sorbent assay technology. Anti-MCP-1 polyclonal antibody was pre-coated onto 96-well plates. And the biotin conjugated anti-MCP-1 polyclonal antibody was used as detection antibodies. The standards test samples and biotin conjugated detection antibody were added - the wells subsequently and wash with wash buffer. Avidin-Biotin-Peroxidase Complex was added and unbound conjugates were washed away with wash buffer. TMB substrates were used - visualize HRP enzymatic reaction. TMB was catalyzed by HRP - produce a blue color product that changed into yellow after adding acidic stop solution. The density of yellow is proportional - the MCP-1 amount of sample captured in plate. Read the O.D. absorbance at 450 nm in a microplate reader and then the concentration of MCP-1 can be calculated.

Plattentyp: Pre-coated

Aufbereitung der Reagenzien:

1. Before the experiment, centrifuge each kit component for several minutes to bring down all reagents to the bottom of tubes. 2. It is recommend to measure each standard and sample in duplicate. 3. Do NOT let the plate completely dry at any time! Since the dry condition can inactivate the biological material on the plate. 4. Do not reuse pipette tips and tubes to avoid cross contamination. 5. Do not use the expired components and the components from different batches. 6. To avoid the marginal effect of plate incubation for temperature differences (the marginal wells always get stronger reaction), it is recommend to equilibrate the ABC working solution and TMB substrate for at least 30 min at room temperature (37 °C ) before adding to wells.The TMB substrate (Kit Component 8) is colorless and transparent before use, if not, please contact us for replacement.

Aufbereitung der Proben:

Preparation of sample and reagents 1. Sample Isolate the test samples soon after collecting, then, analyze immediately (within 2 hours). Or aliquot and store at -20 °C for long term. Avoid multiple freeze-thaw cycles.
Tissue lysate, body fluids and cell culture supernatants: Centrifuge to remove precipitate, analyze immediately or aliquot and store at -20 °C .
Serum: Coagulate the serum at room temperature (about 4 hours). Centrifuge at approximately 2000 × g for 20 min. Analyze the serum immediately or aliquot and store at -20 °C .
Plasma: Collect plasma with EDTA as the anticoagulant. Centrifuge for 20 min at 2000 x g within 30 min of collection. Analyze immediately or aliquot and store frozen at -20 °C. Heparin and citrate can not be used as anticoagulant. Note: 1. Coagulate blood samples completely, then, centrifuge, and avoid hemolysis and particle. 2. NaN3 can not be used as test sample preservative, since it is the inhibitor for HRP.

Beschränkungen: Nur für Forschungszwecke einsetzbar

Handhabung

Konservierungsmittel: Sodium azide, Thimerosal (Merthiolate)

Lagerung: 4 °C,-20 °C

Informationen zur Lagerung: Store at 2-8°C for 6 months, or at -20°C for 12 months.

Haltbarkeit: 12 months

Details zu CCL2

Target: CCL2 (Chemokine (C-C Motif) Ligand 2 (CCL2))

Andere Bezeichnung: MCP-1 / CCL2

Hintergrund: Monocyte chemotactic protein-1(MCP-1), known as chemmkine(C-C motif) ligand 2(CCL2) or small inducible cytokine A2, is a small cytokine belonging to the CC chemokine family. The CCL2 gene is located on chromosome 17 (17q11.2-q21.1) in humans, and spans 1,927 bases. It has three exons and two introns, and produced as a protein precursor containing signal peptide of 23 amino acids and a mature peptide of 76 amino acids. The predicted weight is 11.025 kiloDaltons (kDa). MCP-1 has been implicated in the pathogenesis of diseases characterized by monocytic infiltrates, like psoriasis, rheumatoid arthritis and atherosclerosis. It induces amylin expression through ERK1/ ERK2/ JNK-AP1 and NF-țB related signaling pathways independent of CCR2. Amylin upregulation by CCL2 may contribute to elevation of plasma amylin in obesity and insulin resistance.

Pathways: Cellular Response to Molecule of Bacterial Origin, Positive Regulation of Immune Effector Process, ER-Nucleus Signaling, Unfolded Protein Response, Phosphorylierungen bei SARS-CoV-2 Infektion