Human Chemokine (C-C motif) Ligand 5, CCL5 ELISA Kit (CKERS-CCL5-056H)

Regulatory status: For research use only, not for use in diagnostic procedures.

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Size
15plates, 45plates
Species Reactivity
Human
Intended Use
Human CCL5 ELISA development kit contains the key components required for the quantitative measurement of natural and/or recombinant CCL5 in a sandwich ELISA format within the range of 32-3,000 pg/mL. Using the ELISA protocol described below; kit provides sufficient reagents to assay CCL5 in approximately 1500 ELISA plate wells.
Contents of Kit
1. PBS: Dilute 10xPBS to 1xPBS, pH 7.2 in sterile water.
2. Wash Buffer: 0.05% Tween-20 in PBS
3. Block Buffer: 1.0% BSA in PBS
4. Diluent: 1.0% BSA in PBS
5. ELISA microplates (Thermo Fisher Cat. # 456529);
6. BSA (Sigma Cat. # A-7030);
7. Stop Solution 2 M Sulfuric Acid (Sigma Cat. # 339741);
8. Dulbecco's PBS [10x] (Gibco BRL Cat. # 14200-075).
Storage
Unopened Kit Store at 2-8°C. Do not use past kit expiration date. For more detailed information, please download the document on our website.
General Description
This gene is one of several CC cytokine genes clustered on the q-arm of chromosome 17. Cytokines are a family of secreted proteins involved in immunoregulatory and inflammatory processes. The CC cytokines are proteins characterized by two adjacent cysteines. The cytokine encoded by this gene functions as a chemoattractant for blood monocytes, memory T helper cells and eosinophils. It causes the release of histamine from basophils and activates eosinophils. This cytokine is one of the major HIV-suppressive factors produced by CD8+ cells.
Standard Curve
Reconstitution And Storage
Capture Antibody: 180 μg of antigen-affinity purified goat anti-CCL5. Centrifuge vial prior to opening. Reconstitute in 0.5 mL PBS for a concentration of 360 μg/mL. Following reconstitution the Capture antibodies may be stored at 2-8°C for up to 6 months. For long term storage it is recommended to aliquot into working volumes and store at -70°C in a manual defrost freezer.
Detection Antibody: 3.6 μg of biotinylated antigen-affinity purified goat anti-CCL5. Centrifuge vial prior to opening. Reconstitute in 0.5 mL sterile 1.0% BSA in PBS for a concentration of 7.2 μg/mL. Following reconstitution the Detection antibodies may be stored at 2-8°C for up to 6 months. For long term storage it is recommended to aliquot into working volumes and store at -70°C in a manual defrost freezer.
Human RANTES Standard: 60 ng of recombinant RANTES. Centrifuge vial prior to opening. Reconstitute in 0.5 mL sterile 1.0% BSA in PBS for a concentration of 120 ng/mL. The Standard may be stored at 2-8°C for one month or aliquoted and stored at -70°C for up to three months in a manual defrost freezer.
Avidin-HRP Conjugate: 40 μL vial. Upon receipt, UltraAvidin-HRP conjugate should be stored at 2-8°C, DO NOT FREEZE.
TMB Liquid Substrate: Aspirate and wash plate 4 times. Add 100 μL of TMB HRPO Microwell Substrate Standard Kinetic One Component "Ready Use" to each well. Incubate at room temperature and monitor color development with an ELISA plate reader at 450 nm with wavelength correction set at 540 nm or 570 nm. The reaction may be stopped after 15-20 minutes by adding 100 μL of 2 M sulfuric acid to each well.

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References


Circulating levels of chemokines in patients with psoriasis vulgaris and their association with disease severity: A case-control study from North India

INDIAN JOURNAL OF DERMATOLOGY VENEREOLOGY & LEPROLOGY

Authors: Joshi, Neha; Narang, Tarun; Dogra, Sunil; Chhabra, Seema

Background: Psoriasis is a chronic inflammatory skin disease characterized by hyperproliferation and incomplete differentiation of epidermis, and accumulation of neutrophils and proinflammatory T cells in epidermis and dermis. Chemokines are believed to be the main players mediating the chemotaxis of leucocytes to the lesional site. Previous studies have established the role of various chemokine ligands and receptors at the lesional site in psoriasis. Aims: In this study, we have compared the serum levels of various chemokines, namely, inducible protein-10 (IP-10) (CXCL10), MCP-1 (CCL-2), monokine induced by gamma interferon (MIG) (CXCL-9), RANTES (CCL5), interleukin (IL)-8, and eotaxin in patients with chronic plaque psoriasis with that of healthy controls. We also studied whether the chemokine levels varied within different patient groups based on various clinical and demographic parameters, and if any of these chemokines correlated with disease activity. Methods: We studied 40 patients with chronic plaque psoriasis from a single center. Their clinical and demographic details were recorded in predesigned prforma. Patients with unstable forms of psoriasis like guttate, erythrodermic, or pustular psoriasis were excluded. The serum chemokine levels were measured by flow cytometry-based bead array set system. The serum levels of the patients were compared with that of 25 healthy controls. A subgroup analysis was also done to study the correlation of chemokine levels with age, sex, duration, and severity of disease. Results: We observed a significant decrease in serum level of all these chemokines in patients, when compared with that of healthy controls. We also found that MIG levels showed a positive correlation with disease severity based on Psoriasis Area and Severity Index. Limitations: The major limitation of the study is lack of data on the lesional chemokine levels compared to serum chemokines. Conclusion: The inflammatory process in psoriasis is orchestrated through chemokines. MIG is a potential serum biomarker for assessing disease severity.

Chemokine gene expression influences metastasis and survival time of female dogs with mammary carcinoma

VETERINARY IMMUNOLOGY AND IMMUNOPATHOLOGY

Authors: Ariyarathna, Harsha; Thomson, Neroli; Aberdein, Danielle; Munday, John S.

Chemokines are signaling proteins secreted by immune cells which regulate leukocyte trafficking. The aberrant expression of chemokines and their receptors by neoplastic cells influences the behaviour of many human cancers. This study evaluated gene-expression of the chemokines: CCL5, CXCL10, CXCL12 and the chemokine receptors: CXCR3, CXCR4, CXCR7, CCR4, CCR9 in 41 histologically-malignant, outcome-known, canine mammary tumours. These chemokines and chemokine receptors were selected as all were previously shown to influence the behaviour of human breast cancers. The expression of chemokines CCL5 and CXCL12 were significantly higher in tumours which subsequently metastasised than tumours that did not metastasise (p < 0.05). Increased expression of these chemokines was also correlated with shorter survival times of the dogs (CCL5: r(s) = -0.40, p = 0.02, CXCL12: r(s) = -0.40, p = 0.03) while CCL5 was independently prognostic of survival times (p = 0.026). A significantly higher proportion of tumours that subsequently metastasised expressed CXCR3 (p = 0.037), CXCR4 (p = 0.026), CXCR7 (p = 0.025) and CCR9 (p = 0.039) receptors while the survival times of the dogs with tumours that expressed CXCR4 (p = 0.045) and CCR9 (p = 0.039) receptors were significantly shorter than dogs with tumours that did not express these receptors. Chemokine and chemokine receptor gene-expression has not been previously correlated with disease outcome of canine mammary tumours. These findings indicate that altered expression of chemokines and their receptors influences the behaviour of canine mammary tumours suggesting a potential role of them as prognostic markers or therapeutic targets.

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