Human CCL5/RANTES ELISA Kit

Biofargo Verified independent store

→

{ "@context": "https://schema.org", "@type": "Product", "name": "Human CCL5/RANTES ELISA Kit", "sku": "EK1129", "mpn": "EK1129", "description": "A quantitative sandwich ELISA kit f or determining human Chemokine (C-C motif) Ligand 5, also known as CCL5 or RANTES, concentrations in serum, plasma and cell culture supernatants.", "category": "Human Cytokine ELISA Kit", "additionalProperty": [ { "@type": "PropertyValue", "name": "Target", "value": "Human CCL5/RANTES" }, { "@type": "PropertyValue", "name": "Species", "value": "Human" }, { "@type": "PropertyValue", "name": "Assay Type", "value": "Quantitative sandwich ELISA" }, { "@type": "PropertyValue", "name": "Sample Types", "value": "Serum, plasma and cell culture supernatant" }, { "@type": "PropertyValue", "name": "Sensitivity", "value": "0.53 pg/mL" }, { "@type": "PropertyValue", "name": "Standard Range", "value": "15.63–1,000 pg/mL" }, { "@type": "PropertyValue", "name": "Assay Time", "value": "3.5 hours" }, { "@type": "PropertyValue", "name": "Detection", "value": "HRP-TMB colorimetric detection at 450 nm" }, { "@type": "PropertyValue", "name": "Available Sizes", "value": "48T and 96T" }, { "@type": "PropertyValue", "name": "Storage", "value": "2–8°C for unopened kits" } ] } { "@context": "https://schema.org", "@type": "FAQPage", "mainEntity": [ { "@type": "Question", "name": "What does the Human CCL5/RANTES ELISA Kit measure?", "acceptedAnswer": { "@type": "Answer", "text": "The kit is intended for the quantitative determination of human Chemokine (C-C motif) Ligand 5, also known as CCL5 or RANTES." } }, { "@type": "Question", "name": "Which sample types can be tested with this Human CCL5 ELISA Kit?", "acceptedAnswer": { "@type": "Answer", "text": "The kit has been tested with human serum, plasma and cell culture supernatants. Other biological samples may be suitable, but sample dilution or further evaluation may be required." } }, { "@type": "Question", "name": "Which plasma anticoagulants are compatible with the assay?", "acceptedAnswer": { "@type": "Answer", "text": "Plasma may be collected using EDTA, citrate or heparin as the anticoagulant." } }, { "@type": "Question", "name": "What is the sensitivity of the Human CCL5/RANTES ELISA Kit?", "acceptedAnswer": { "@type": "Answer", "text": "The stated sensitivity is 0.53 pg/mL." } }, { "@type": "Question", "name": "What is the standard range of the CCL5/RANTES ELISA assay?", "acceptedAnswer": { "@type": "Answer", "text": "The standard range is 15.63 to 1,000 pg/mL." } }, { "@type": "Question", "name": "How should normal serum and plasma samples be diluted?", "acceptedAnswer": { "@type": "Answer", "text": "Normal serum and plasma samples require a 100-fold dilution. The manual suggests combining 5 microliters of sample with 495 microliters of 1X Assay Buffer." } }, { "@type": "Question", "name": "What wavelength is required to read the ELISA plate?", "acceptedAnswer": { "@type": "Answer", "text": "Measure absorbance at 450 nm. When available, use a correction wavelength of 570 nm or 630 nm." } }, { "@type": "Question", "name": "What assay method is used?", "acceptedAnswer": { "@type": "Answer", "text": "The kit uses a quantitative sandwich enzyme immunoassay with a pre-coated capture antibody, a biotin-linked detection antibody, streptavidin-HRP and TMB colorimetric substrate." } }, { "@type": "Question", "name": "How long does the CCL5/RANTES ELISA assay take?", "acceptedAnswer": { "@type": "Answer", "text": "The stated assay time is approximately 3.5 hours." } }, { "@type": "Question", "name": "How should samples be stored before testing?", "acceptedAnswer": { "@type": "Answer", "text": "Samples tested within 24 hours may be stored at 2–8°C. For longer storage, aliquot samples and store them at or below -20°C while avoiding repeated freeze-thaw cycles." } }, { "@type": "Question", "name": "How should the ELISA kit be stored?", "acceptedAnswer": { "@type": "Answer", "text": "Store the unopened kit at 2–8°C. After opening, the reconstituted standard may be stored at -20°C for approximately one month and should be discarded after a single-use reconstitution. Other components may be stored at 2–8°C for approximately one month." } }, { "@type": "Question", "name": "Is this Human CCL5/RANTES ELISA Kit intended for diagnostic use?", "acceptedAnswer": { "@type": "Answer", "text": "No. The kit is for research use only and is not intended for use in diagnostic procedures." } } ] } Description Accurate measurement of human CCL5, also known as RANTES, requires an assay that can quantify low concentrations across commonly used biological sample types while supporting consistent plate-based workflows. The Human CCL5/RANTES ELISA Kit is designed for the quantitative determination of human CCL5/RANTES concentrations in serum, plasma and cell culture supernatants. This quantitative sandwich ELISA uses a microplate pre-coated with a monoclonal antibody specific for human CCL5. Bound CCL5 is detected with a biotin-linked detection antibody, followed by streptavidin-HRP and TMB color development. The resulting color intensity is proportional to the amount of human CCL5/RANTES present in the sample. The assay provides a sensitivity of 0.53 pg/mL and a standard range of 15.63–1,000 pg/mL. Absorbance is measured at 450 nm using a standard microplate reader, with wavelength correction at 570 nm or 630 nm when available. For research use only. Not for use in diagnostic procedures. Quick Details Product Name Human CCL5/RANTES ELISA Kit Catalog Number EK1129 Target Human Chemokine (C-C Motif) Ligand 5 (CCL5/RANTES) Assay Format Quantitative Sandwich ELISA Sample Types Serum, Plasma, and Cell Culture Supernatant Sensitivity 0.53 pg/mL Standard Range 15.63–1,000 pg/mL Assay Time Approximately 3.5 hours Detection HRP-TMB Colorimetric Detection Readout 450 nm with Optional 570 nm or 630 nm Correction Available Sizes 48 Tests (48T) and 96 Tests (96T) Specifications Name Human CCL5/RANTES ELISA Kit Catalog Number EK1129 HGNC ID 10632 Species Human Target Names CCL5, RANTES, Chemokine (C-C Motif) Ligand 5 Assay Type Quantitative Sandwich Enzyme Immunoassay Assay Time 3.5 hours Sample Type Serum, Plasma, Cell Culture Supernatant, and Other Potentially Suitable Biological Samples Validated Sample Types Serum, Plasma, and Cell Culture Supernatant Compatible Plasma Types EDTA Plasma, Citrate Plasma, and Heparin Plasma Sample Amount 20 µL of Prediluted Serum or Plasma per Well Cell Culture Supernatant Volume 100 µL per Well Sensitivity 0.53 pg/mL Standard Range 15.63–1,000 pg/mL Recovery 84%–117% Average Recovery 97% Intra-Assay Precision 6.9%–7.8% Inter-Assay Precision 5.5%–8.3% Plate Type Detachable 96-Well Plate Platform ELISA Detection System Biotin-Linked Detection Antibody, Streptavidin-HRP, and TMB Substrate Measurement Wavelength 450 nm Correction Wavelength 570 nm or 630 nm Kit Storage 2–8 °C Shipping Condition 4 °C with Blue Ice Research Use For Research Use Only; Not for Use in Diagnostic Procedures Features Quantitative measurement of human CCL5/RANTES concentrations. Quantitative sandwich ELISA assay format. Monoclonal anti-human CCL5 capture antibody pre-coated on the microplate. Biotin-linked CCL5 detection antibody with streptavidin-HRP detection. HRP-TMB colorimetric readout at 450 nm. Suitable for human serum, plasma and cell culture supernatants. Compatible with EDTA, citrate and heparin plasma. 0.53 pg/mL stated sensitivity. 15.63–1,000 pg/mL standard range. Available in 48T and 96T kit sizes. Applications The Human CCL5/RANTES ELISA Kit is intended for quantitative CCL5 measurement in research samples. Supported applications include: Quantification of human CCL5/RANTES in serum samples. Measurement of human RANTES concentrations in EDTA, citrate or heparin plasma. Detection of secreted CCL5 in cell culture supernatants. Human chemokine profiling studies. Research involving CCL5-associated inflammatory responses. Studies of leukocyte recruitment to inflammatory sites. Research involving the chemotactic activity of CCL5 toward T cells, eosinophils and basophils. Studies involving cytokine-associated natural killer cell activation. Research examining CCL5 released by CD8-positive T cells. This product is intended exclusively for research use. It is not intended for disease diagnosis, patient screening or therapeutic decision-making. Principle of the Assay This assay employs a quantitative sandwich enzyme immunoassay technique. A monoclonal antibody specific for human CCL5 is pre-coated onto the microplate. Standards and samples are added to the coated wells. A biotin-linked detection antibody specific for human CCL5 is added. Human CCL5 present in the sample is bound between the immobilized capture antibody and the detection antibody. Unbound substances are removed by washing. Streptavidin-HRP is added and binds to the biotin-linked detection antibody. After another wash, TMB substrate is added. Color develops in proportion to the amount of CCL5 bound in the well. Stop Solution is added, and absorbance is measured at 450 nm. Standard Curve A standard curve must be generated during each assay. The standard series described in the product manual includes the following concentrations: Standard Human CCL5/RANTES Concentration Standard 1 1,000 pg/mL Standard 2 500 pg/mL Standard 3 250 pg/mL Standard 4 125 pg/mL Standard 5 62.50 pg/mL Standard 6 31.25 pg/mL Standard 7 15.63 pg/mL Zero Standard 0 pg/mL The standard curve data supplied in the manual are provided for demonstration only. A new standard curve must be run with each assay. Sample Collection and Preparation Cell Culture Supernatants Remove particulates by centrifugation. Test freshly prepared samples immediately or divide them into aliquots and store at or below -20°C. Avoid repeated freeze-thaw cycles. Cell culture supernatants may be added at 100 µL per well. Samples with high CCL5/RANTES concentrations may be diluted using the corresponding cell culture medium. Serum Collect blood using a serum separator tube and allow it to clot for 30 minutes. Centrifuge for 10 minutes at 1,000 × g. Remove the serum and test it immediately or divide it into aliquots and store at or below -20°C. Normal serum samples require a 100-fold dilution. The suggested dilution is: 5 µL serum sample + 495 µL 1X Assay Buffer Plasma Collect plasma using EDTA, citrate or heparin as the anticoagulant. Centrifuge at 1,000 × g within 30 minutes of collection. Test freshly prepared plasma immediately or divide it into aliquots and store at or below -20°C. Normal plasma samples require a 100-fold dilution. The suggested dilution is: 5 µL plasma sample + 495 µL 1X Assay Buffer General Sample Handling Clarify samples containing visible precipitates before testing. Do not use grossly hemolyzed or lipemic specimens. Samples tested within 24 hours may be stored at 2–8°C. For longer storage, aliquot samples and store at -20°C. Avoid repeated freeze-thaw cycles. Dilution with 1X Assay Buffer may be necessary for samples with high CCL5/RANTES concentrations. Assay Workflow Bring all kit reagents and samples to room temperature before use. Prepare the standards, samples, detection antibody, streptavidin-HRP, Assay Buffer and Washing Buffer. Wash the required microplate wells once using 300 µL of 1X Washing Buffer per well. Add 100 µL of each two-fold diluted standard to the standard wells. Add 100 µL of Assay Buffer or culture medium to the blank well. For serum or plasma, add 80 µL of 1X Assay Buffer and 20 µL of prediluted sample per well. For cell culture supernatants, add 100 µL of sample per well. Add 50 µL of diluted detection antibody to each well. Seal the plate and incubate for 2 hours at 25°C ± 3°C on a microplate shaker at 300 rpm. Wash the wells six times using 300 µL of 1X Washing Buffer per wash. Add 100 µL of diluted streptavidin-HRP to each well. Incubate for 45 minutes at 25°C ± 3°C on a microplate shaker at 300 rpm. Wash the wells six times. Add 100 µL of TMB substrate to each well. Incubate for 5–30 minutes at room temperature while protecting the plate from light. Add 100 µL of Stop Solution to each well. Measure optical density within 30 minutes at 450 nm, with correction at 570 nm or 630 nm when available. Kit Components Component 48T Kit 96T Kit Anti-CCL5 Monoclonal Antibody-Coated Microplate 48 Tests 96 Tests Freeze-Dried Human CCL5/RANTES Standard 1 Vial 2 Vials CCL5/RANTES Detection Antibody 1 Vial 1 Vial Streptavidin-HRP 1 Vial 1 Vial Assay Buffer, 10X 10 mL 10 mL TMB Substrate 6 mL 11 mL Stop Solution 11 mL 11 mL Washing Buffer, 20X 50 mL 50 mL Adhesive Films 6 6 Components from reagent kits with different batch numbers must not be used interchangeably. Additional Equipment Required Microplate reader capable of measuring absorbance at 450 nm. Optional wavelength correction at 570 nm or 630 nm. Single-channel and multichannel micropipettes with disposable tips. 50–300 µL adjustable multichannel micropipette. Multichannel micropipette reservoir. Polypropylene tubes for standard and sample dilution. Deionized or distilled water. Laboratory vessels required for reagent preparation. Microplate shaker capable of operating at 300 rpm. Storage and Shipping Unopened kit: Store at 2–8°C and refer to the product label for the expiration date. Opened microplate: Return unused strips to the aluminum foil bag, reseal the bag and store at 2–8°C for approximately one month. Reconstituted standard: Store at -20°C for approximately one month and discard after single-use reconstitution. Other opened components: Store at 2–8°C for approximately one month. Prepared 1X Washing Buffer: Store at 2–25°C for up to 30 days. Prepared 1X Assay Buffer: Store at 2–8°C for up to 30 days. Shipping condition: Transport at approximately 4°C with blue ice. Documents Human CCL5/RANTES ELISA Kit User Manual Frequently Asked Questions What does the Human CCL5/RANTES ELISA Kit measure? The kit is intended for the quantitative determination of human Chemokine (C-C motif) Ligand 5, also known as CCL5 or RANTES. Which sample types can be tested with this Human CCL5 ELISA Kit? The kit has been tested with human serum, plasma and cell culture supernatants. Other biological samples may be suitable, but sample dilution or further evaluation may be required. Which plasma anticoagulants are compatible with the assay? Plasma may be collected using EDTA, citrate or heparin as the anticoagulant. Samples should be centrifuged at 1,000 × g within 30 minutes of collection. What is the sensitivity of the Human CCL5/RANTES ELISA Kit? The stated sensitivity is 0.53 pg/mL. What is the standard range of the CCL5/RANTES ELISA assay? The standard range is 15.63–1,000 pg/mL. A standard curve must be prepared and run with every assay. How should normal serum and plasma samples be diluted? Normal serum and plasma samples require a 100-fold dilution. The suggested preparation is 5 µL of sample combined with 495 µL of 1X Assay Buffer. How much serum or plasma sample is added to each well? Add 80 µL of 1X Assay Buffer and 20 µL of prediluted serum or plasma sample to each sample well. How much cell culture supernatant is required? Add 100 µL of cell culture supernatant to each sample well. Samples with high CCL5/RANTES concentrations may be diluted with the corresponding cell culture medium. What wavelength should be used to read the ELISA plate? Measure absorbance at 450 nm. When available, use a correction wavelength of 570 nm or 630 nm. Measurements should be completed within 30 minutes after adding Stop Solution. What assay method is used? The kit uses a quantitative sandwich enzyme immunoassay. Human CCL5 is captured by a pre-coated monoclonal antibody and detected using a biotin-linked detection antibody, streptavidin-HRP and TMB substrate. How long does the Human RANTES ELISA assay take? The stated assay time is approximately 3.5 hours. The workflow includes a 2-hour sample and detection antibody incubation, a 45-minute streptavidin-HRP incubation and a 5–30-minute substrate incubation. How should biological samples be stored before testing? Samples tested within 24 hours may be stored at 2–8°C. For longer storage, divide samples into aliquots and store them at or below -20°C. Avoid repeated freeze-thaw cycles. Can visibly precipitated, hemolyzed or lipemic samples be tested? Samples containing visible precipitates must be clarified before use. Grossly hemolyzed or lipemic specimens should not be used. How should the Human CCL5/RANTES ELISA Kit be stored after opening? Unused microplate strips and most other components may be stored at 2–8°C for approximately one month. The reconstituted standard may be stored at -20°C for approximately one month and should be discarded after single-use reconstitution. Is this Human CCL5/RANTES ELISA Kit intended for diagnostic use? No. The kit is for research use only and is not intended for use in diagnostic procedures. For research use only. Not for use in diagnostic procedures. Show more