CCL8 ELISA Kit

Produktnummer ABIN625040

Produktdetails CCL8 ELISA Kit

Target: CCL8 (Chemokine (C-C Motif) Ligand 8 (CCL8))

Reaktivität: Human

Nachweismethode: Colorimetric

Methodentyp: Sandwich ELISA

Detektionsbereich: 1.5-200 pg/mL

Untere Nachweisgrenze: 1.5 pg/mL

Applikation: ELISA

Verwendungszweck: Human MCP-2 (CCL8) 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 any of the cytokines tested: Human Angiogenin, BDNF, BLC, ENA-78, FGF-4, IL-1 alpha, IL-1 beta, IL-2, IL-3, IL-4, IL-5, IL-7, IL-8, IL-9, IL-10, IL-11, IL-12 p70, IL-12 p40, IL-13, IL-15, I-309, IP-10, G-CSF, GM-CSF, IFN-gamma, Leptin, MCP-1, MCP-3, MDC, MIP-1 alpha, MIP-1 beta, MIP-1 delta, PARC, PDGF, RANTES, SCF, TARC, TGF-beta, TIMP-1, TIMP-2, TNF-alpha, TNF-beta, TPO, VEGF.

Sensitivität: 1.5 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 samples2 - 5 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) is used for dilution of serum/plasma samples, and Assay Diluent B (Item E) is used for dilution of culture supernatants and urine. 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 and urine) into Item C vial to prepare a 50 ng/ml standard. Dissolve the powder thoroughly by a gentle mix. Add 4 µl MCP-2 standard from the vial of Item C, into a tube with 996 µl Assay Diluent A or 1x Assay Diluent B to prepare a 200 pg/ml stock standard solution. Pipette 300 µl Assay Diluent A or 1x Assay Diluent B into each tube. Use the stock standard solution to produce a dilution series (shown below). Mix each tube thoroughly before the next transfer. Assay Diluent A or 1x Assay Diluent B serves as the zero standard (0 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 5,000-fold with 1x Assay Diluent B. For example: Briefly spin the vial (Item G) and pipette up and down to mix gently . Add 3 µl of HRP-Streptavidin concentrate into a tube with 15 ml 1x Assay Diluent B to prepare 5,000 fold diluted HRP- Streptavidin solution.

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 µl) 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 3. 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 3. 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.

Beschränkungen: Nur für Forschungszwecke einsetzbar

Handhabung

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 CCL8 ELISA Kit (ABIN625040)

Petrosino, Puigdemont, Della Valle, Fusco, Verde, Allarà, Aveta, Orlando, Di Marzo: "Adelmidrol increases the endogenous concentrations of palmitoylethanolamide in canine keratinocytes and down-regulates an inflammatory reaction in an in vitro model of contact allergic dermatitis." in: Veterinary journal (London, England : 1997), Vol. 207, pp. 85-91, (2016) (PubMed).

Ahrens, Schulz, Bellach, Niggemann, Beyer: "Chemokine levels in serum of children with atopic dermatitis with regard to severity and sensitization status." in: Pediatric allergy and immunology : official publication of the European Society of Pediatric Allergy and Immunology, Vol. 26, Issue 7, pp. 634-40, (2015) (PubMed).

Poole, Avdic, Hodkinson, Jackson, Wills, Slobedman, Sinclair et al.: "Latency-associated viral interleukin-10 (IL-10) encoded by human cytomegalovirus modulates cellular IL-10 and CCL8 Secretion during latent infection through changes in the cellular microRNA ..." in: Journal of virology, Vol. 88, Issue 24, pp. 13947-55, (2014) (PubMed).

Hamilton, Tower, Jones: "Identification of chemokines associated with the recruitment of decidual leukocytes in human labour: potential novel targets for preterm labour." in: PLoS ONE, Vol. 8, Issue 2, pp. e56946, (2013) (PubMed).

Liu, Liu, Chen, Chen, He, Zheng, Yang, Song, Weng, Hu, Fan, Xiao, Kaufmann, Ernst, Ge: "Induction of CCL8/MCP-2 by mycobacteria through the activation of TLR2/PI3K/Akt signaling pathway." in: PLoS ONE, Vol. 8, Issue 2, pp. e56815, (2013) (PubMed).

Chen, Huang, Kuo, Lo, Lee, Wang: "Inhibition of glutaminyl cyclase attenuates cell migration modulated by monocyte chemoattractant proteins." in: The Biochemical journal, Vol. 442, Issue 2, pp. 403-12, (2012) (PubMed).

Wang, Prakash, Nguyen, Davis-Dusenbery, Hill, Layne, Hata, Lagna: "Bone morphogenetic protein signaling in vascular disease: anti-inflammatory action through myocardin-related transcription factor A." in: The Journal of biological chemistry, Vol. 287, Issue 33, pp. 28067-77, (2012) (PubMed).

Yi, Elson, Mitsuhashi, Jacobs, Hollovary, Budd, Spiro, Triozzi, Borden: "Phosphatase inhibitor, sodium stibogluconate, in combination with interferon (IFN) alpha 2b: phase I trials to identify pharmacodynamic and clinical effects." in: Oncotarget, Vol. 2, Issue 12, pp. 1155-64, (2012) (PubMed).

Borden, Jacobs, Hollovary, Rybicki, Elson, Olencki, Triozzi: "Gene regulatory and clinical effects of interferon β in patients with metastatic melanoma: a phase II trial." in: Journal of interferon & cytokine research : the official journal of the International Society for Interferon and Cytokine Research, Vol. 31, Issue 5, pp. 433-40, (2011) (PubMed).

Details zu CCL8

Target: CCL8 (Chemokine (C-C Motif) Ligand 8 (CCL8))

Andere Bezeichnung: MCP-2 (CCL8 Produkte)

Synonyme: HC14 ELISA Kit, MCP-2 ELISA Kit, MCP2 ELISA Kit, SCYA10 ELISA Kit, SCYA8 ELISA Kit, 1810063B20Rik ELISA Kit, AB023418 ELISA Kit, Mcp2 ELISA Kit, Scya8 ELISA Kit, C-C motif chemokine ligand 8 ELISA Kit, chemokine (C-C motif) ligand 8 ELISA Kit, C-C motif chemokine 8 ELISA Kit, monocyte chemoattractant protein-2 precursor ELISA Kit, CCL8 ELISA Kit, Ccl8 ELISA Kit, LOC101119572 ELISA Kit, MCP-2 ELISA Kit

Hintergrund: Monocyte chemotactic protein-2 (MCP-2) is found in the conditioned medium of human osteosarcoma cell line MG-63 that also secretes some other chemotactic factors including IL-8 and MCP-1. MCP-2 is related closely to MCP-1 and MCP-3. MCP-2 is a member of the family of CC-Chemokines and specifically attracts monocytes, but not neutrophils, in vitro. Intradermal injection into rabbits causes the selective recruitment of monocytes at the site of injection. The factor does not possess suppressive activity against immature subsets of myeloid progenitors stimulated to proliferate by multiple growth factors. The Human MCP-2 ELISA (Enzyme-Linked Immunosorbent Assay) kit is an in vitro enzyme-linked immunosorbent assay for the quantitative measurement of human MCP-2 in serum, plasma, cell culture supernatants and urine. This assay employs an antibody specific for human MCP-2 coated on a 96-well plate. Standards and samples are pipetted into the wells and MCP-2 present in a sample is bound to the wells by the immobilized antibody. The wells are washed and biotinylated anti-human MCP-2 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-2 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: 6355

UniProt: P80075