CCL5 Kit ELISA

N° du produit ABIN1979657

Détail du produit CCL5 Kit ELISA

Antigène: CCL5 (Chemokine (C-C Motif) Ligand 5 (CCL5))

Reactivité: Humain

Méthode de détection: Colorimetric

Type de méthode: Sandwich ELISA

Gamme de detection: 3-2000 pg/mL

Seuil minimal de détection: 3 pg/mL

Application: ELISA

Fonction: Human RANTES (CCL5) 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: 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-2, MCP-3, MDC, MIP-1 alpha, MIP-1 beta, MIP-1 delta, PARC, PDGF, SCF, TARC, TGF-beta, TIMP-1, TIMP-2, TNF-alpha, TNF-beta, TPO, 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 samples2 - 50 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 and urine. Suggested dilution for normal serum/plasma: 2-50 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 and urine) into Item C vial to prepare a 50 ng/mL standard. Dissolve the powder thoroughly by a gentle mix. Add 40 µL RANTES standard from the vial of Item C, into a tube with 960 µL Assay Diluent A or 1x Assay Diluent B to prepare a 2000 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 40 µL standard +960 µL 2000 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 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) before use. HRP-Streptavidin concentrate should be diluted 300-fold with 1x Assay Diluent B. For example: Briefly spin the vial (Item G) and pipette up and down to mix gently . Add 40 µL of HRP-Streptavidin concentrate into a tube with 12 ml 1x Assay Diluent B to prepare a 300-fold diluted HRP-Streptavidin solution (don't store the diluted solution for next day use). 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 Rantes concentration (pg/mL) 1 10 100 1000 10000 O D =4 50 n m 0.001 0.01 0.1 1 10 Assay Diluent B Rantes concentration (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 RANTES is typically less than 3 pg/mL.
Recovery: Recovery was determined by spiking various levels of human RANTES into human serum, plasma and cell culture media. Mean recoveries are as follows: Sample Type Average % Recovery Range ( %) Serum 96.74 85-104 Plasma 94.76 84-103 Cell culture media 102.6 87-108
Linearity: Sample Type Serum Plasma Cell culture media 1:2 Average % of Expected 95 97 98 Range ( %) 85-105 86-105 86-106 1:4 Average % of Expected 97 98 102 Range ( %) 86-104 85-106 89-108
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 CCL5 Kit ELISA (ABIN1979657)

Persidsky, Hill, Zhang, Dykstra, Winfield, Reichenbach, Potula, Mukherjee, Ramirez, Rom: "Dysfunction of brain pericytes in chronic neuroinflammation." dans: Journal of cerebral blood flow and metabolism : official journal of the International Society of Cerebral Blood Flow and Metabolism, Vol. 36, Issue 4, pp. 794-807, (2016) (PubMed).

Khoo, Nadarajan, Shim, Miswan, Zang, Possinger, Elstner: "Pretreatment of BMSCs with TZD solution decreases the proliferation rate of MCF‑7 cells by reducing FGF4 protein expression." dans: Molecular medicine reports, Vol. 13, Issue 4, pp. 3406-14, (2016) (PubMed).

Tuchscherr, Bischoff, Lattar, Noto Llana, Pförtner, Niemann, Geraci, Van de Vyver, Fraunholz, Cheung, Herrmann, Völker, Sordelli, Peters, Löffler: "Sigma Factor SigB Is Crucial to Mediate Staphylococcus aureus Adaptation during Chronic Infections." dans: PLoS pathogens, Vol. 11, Issue 4, pp. e1004870, (2016) (PubMed).

Lipkova, Parenica, Duris, Helanova, Tomandl, Kubkova, Vasku, Goldbergova Pavkova: "Association of circulating levels of RANTES and -403G/A promoter polymorphism to acute heart failure after STEMI and to cardiogenic shock." dans: Clinical and experimental medicine, Vol. 15, Issue 3, pp. 405-14, (2015) (PubMed).

Zemskova, Song, Cen, Cerda-Infante, Montecinos, Kraft: "Regulation of prostate stromal fibroblasts by the PIM1 protein kinase." dans: Cellular signalling, Vol. 27, Issue 1, pp. 135-46, (2015) (PubMed).

Chen, Eksioglu, Carter, Fortenbery, Donatelli, Zhou, Liu, Yang, Gilvary, Djeu, Wei: "Inactivation of DAP12 in PMN inhibits TREM1-mediated activation in rheumatoid arthritis." dans: PLoS ONE, Vol. 10, Issue 2, pp. e0115116, (2015) (PubMed).

Ciaffi, Cavassini, Genne, Delhumeau, Spycher Elbes, Hill, Wandeler, Fehr, Stoeckle, Schmid, Hirschel, Montecucco, Calmy: "Switch to etravirine for HIV-positive patients receiving statin treatment: a prospective study." dans: European journal of clinical investigation, Vol. 45, Issue 7, pp. 720-30, (2015) (PubMed).

Zemer-Wassercug, Haim, Leshem-Lev, Orvin, Vaduganathan, Gutstein, Kadmon, Mager, Kornowski, Lev, Lev: "The effect of dabigatran and rivaroxaban on platelet reactivity and inflammatory markers." dans: Journal of thrombosis and thrombolysis, Vol. 40, Issue 3, pp. 340-6, (2015) (PubMed).

Israelow, Narbus, Sourisseau, Evans: "HepG2 cells mount an effective antiviral interferon-lambda based innate immune response to hepatitis C virus infection." dans: Hepatology (Baltimore, Md.), Vol. 60, Issue 4, pp. 1170-9, (2015) (PubMed).

Parnell, Gatt, Krupa, Nickles, McKay, Schibeci, Batten, Baranzini, Henderson, Barnett, Slee, Vucic, Stewart, Booth: "The autoimmune disease-associated transcription factors EOMES and TBX21 are dysregulated in multiple sclerosis and define a molecular subtype of disease." dans: Clinical immunology (Orlando, Fla.), Vol. 151, Issue 1, pp. 16-24, (2014) (PubMed).

Shangguan, Ti, Krause, Hai, Zhao, Yang, Liu: "Inhibition of TGF-?/Smad signaling by BAMBI blocks differentiation of human mesenchymal stem cells to carcinoma-associated fibroblasts and abolishes their protumor effects." dans: Stem cells (Dayton, Ohio), Vol. 30, Issue 12, pp. 2810-9, (2012) (PubMed).

Romero, Coyle, Hayward: "Structure and magnetism of Sr3Co2O4Cl2--an electronically driven lattice distortion in an oxychloride containing square planar Co(II) centers." dans: Journal of the American Chemical Society, Vol. 134, Issue 38, pp. 15946-52, (2012) (PubMed).

Pavkova Goldbergova, Lipkova, Pavek, Gatterova, Vasku, Soucek, Nemec: "RANTES, MCP-1 chemokines and factors describing rheumatoid arthritis." dans: Molecular immunology, Vol. 52, Issue 3-4, pp. 273-8, (2012) (PubMed).

Keskin, Keskin, Kucukosmanoglu, Ozkars, Gogebakan, Kul, Bayram, Coskun: "Exhaled RANTES and interleukin 4 levels after exercise challenge in children with asthma." dans: Annals of allergy, asthma & immunology : official publication of the American College of Allergy, Asthma, & Immunology, Vol. 109, Issue 5, pp. 303-8, (2012) (PubMed).

Eichbaum, Meyer, Wang, Bischofs, Steinborn, Bruckner, Brodt, Sohn, Eichbaum: "Breast cancer cell-derived cytokines, macrophages and cell adhesion: implications for metastasis." dans: Anticancer research, Vol. 31, Issue 10, pp. 3219-27, (2011) (PubMed).

Wiedermann, Kowald, Reinisch, Kaehler, von Luettichau, Pattison, Huie, Sibley, Nelson, Krensky: "Monocyte haptotaxis induced by the RANTES chemokine." dans: Current biology : CB, Vol. 3, Issue 11, pp. 735-9, (2004) (PubMed).

Brenner, Nakayama, Goebl, Tanaka, Toh-e, Matsumoto: "CDC33 encodes mRNA cap-binding protein eIF-4E of Saccharomyces cerevisiae." dans: Molecular and cellular biology, Vol. 8, Issue 8, pp. 3556-9, (1989) (PubMed).

Watson: "Potassium reabsorption in the proximal tubule of the dog nephron." dans: The Journal of clinical investigation, Vol. 45, Issue 8, pp. 1341-8 (PubMed).

Détail du antigène

Antigène: CCL5 (Chemokine (C-C Motif) Ligand 5 (CCL5))

Autre désignation: RANTES (CCL5 Produits)

Synonymes: Rantes Kit ELISA, Scya5 Kit ELISA, D17S136E Kit ELISA, RANTES Kit ELISA, SCYA5 Kit ELISA, SIS-delta Kit ELISA, SISd Kit ELISA, TCP228 Kit ELISA, eoCP Kit ELISA, MuRantes Kit ELISA, C-C motif chemokine 5-like Kit ELISA, C-C motif chemokine ligand 5 Kit ELISA, chemokine (C-C motif) ligand 5 Kit ELISA, LOC100230935 Kit ELISA, CCL5 Kit ELISA, Ccl5 Kit ELISA

Sujet: RANTES is a protein, which belongs to the family of chemotactic cytokines. It is produced by circulating T-cells and T-cell clones in culture. RANTES activates human basophils from some select basophil donors and causes the release of histamines. It can induce the proliferation and activation of killer cells known as CHAK. RANTES is expressed by human synovial fibroblasts and may participate, therefore, in the ongoing inflammatory process in rheumatoid arthritis. The Human RANTES ELISA (Enzyme-Linked Immunosorbent Assay) kit is an in vitro enzyme-linked immunosorbent assay for the quantitative measurement of human RANTES in serum, plasma, cell culture supernatants and urine. This assay employs an antibody specific for human RANTES coated on a 96-well plate. Standards and samples are pipetted into the wells and RANTES present in a sample is bound to the wells by the immobilized antibody. The wells are washed and biotinylated anti-human RANTES 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 RANTES 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%.

ID gène: 6352

UniProt: P13501

Pathways: Cellular Response to Molecule of Bacterial Origin, Regulation of G-Protein Coupled Receptor Protein Signaling, Smooth Muscle Cell Migration