CCL5 ELISA Kit (Guinea Pig)

Intra-assay Precision (Precision within an assay) : 3 samples with low, middle and high level Regulated On Activation In Normal T-Cell Expressed And Secreted (RANTES) were tested 20 times on one plate, respectively. Inter-assay Precision (Precision between assays) : 3 samples with low, middle and high level Regulated On Activation In Normal T-Cell Expressed And Secreted (RANTES) were tested on 3 different plates, 8 replicates in each plate. CV (%) = SD/meanX100 Intra-Assay: CV<10% Inter-Assay: CV<12%

Chemoattractant for blood monocytes, memory T-helper cells and eosinophils. Causes the release of histamine from basophils and activates eosinophils. May activate several chemokine receptors including CCR1, CCR3, CCR4 and CCR5. May also be an agonist of the G protein-coupled receptor GPR75. Together with GPR75, may play a role in neuron survival through activation of a downstream signaling pathway involving the PI3, Akt and MAP kinases. By activating GPR75 may also play a role in insulin secretion by islet cells.

The test principle applied in this kit is Sandwich enzyme immunoassay. The microtiter plate provided in this kit has been pre-coated with an antibody specific to Regulated On Activation In Normal T-Cell Expressed And Secreted (RANTES) . Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated antibody specific to Regulated On Activation In Normal T-Cell Expressed And Secreted (RANTES) . Next, Avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. After TMB substrate solution is added, only those wells that contain Regulated On Activation In Normal T-Cell Expressed And Secreted (RANTES), biotin-conjugated antibody and enzyme-conjugated Avidin will exhibit a change in color. The enzyme-substrate reaction is terminated by the addition of sulphuric acid solution and the color change is measured spectrophotometrically at a wavelength of 450nm +/- 10nm. The concentration of Regulated On Activation In Normal T-Cell Expressed And Secreted (RANTES) in the samples is then determined by comparing the O.D. of the samples to the standard curve.

Reconstitution & Storage Instructions Western Blotting/Immunoblotting (WB/IB) Protocol Immunohistochemistry (IHC) Protocol Immunocytochemistry (ICC) Protocol Enzyme-Linked ImmunoSorbent Assay (ELISA) Protocol Blocking Peptide Competition Protocol (BPCP) Immunoprecipitation (IP) Protocol Antibody Array (AA) Protocol Reconstitution & Storage Instructions Reconstitution & Storage Instructions Western Blotting/Immunoblotting (WB/IB) Protocol Western Blotting/Immunoblotting (WB/IB) Protocol Immunohistochemistry (IHC) Protocol Immunohistochemistry (IHC) Protocol Immunocytochemistry (ICC) Protocol Immunocytochemistry (ICC) Protocol Enzyme-Linked ImmunoSorbent Assay (ELISA) Protocol Enzyme-Linked ImmunoSorbent Assay (ELISA) Protocol Blocking Peptide Competition Protocol (BPCP) Blocking Peptide Competition Protocol (BPCP) Immunoprecipitation (IP) Protocol Immunoprecipitation (IP) Protocol Antibody Array (AA) Protocol Antibody Array (AA) Protocol

Matrix Recovery Range (%) Average (%) serum (n=5) 86-101 94 EDTA plasma (n=5) 79-92 85 heparin plasma (n=5) 96-105 102 Matrix Recovery Range (%) Average (%) Matrix Recovery Range (%) Average (%) serum (n=5) 86-101 94 serum (n=5) 86-101 94 EDTA plasma (n=5) 79-92 85 EDTA plasma (n=5) 79-92 85 heparin plasma (n=5) 96-105 102 heparin plasma (n=5) 96-105 102

Component Amount Anti-CCL5 Microplate 96 Wells (12 x 8 Well Strips) CCL5 Lyophilized Standard 2 x 10 ng 100X Biotinylated CCL5 Detector Antibody 120 uL Avidin/HRP Conjugate 120 uL Standard Diluent 1 x 20 mL Detector Antibody Diluent 1 x 12 mL Conjugate Diluent 1 x 12 mL 30X Wash Buffer 1 x 20 mL TMB Substrate 1 x 9 mL Stop Solution 1 x 6 mL Component Amount Component Amount Anti-CCL5 Microplate 96 Wells (12 x 8 Well Strips) Anti-CCL5 Microplate 96 Wells (12 x 8 Well Strips) CCL5 Lyophilized Standard 2 x 10 ng CCL5 Lyophilized Standard 2 x 10 ng 100X Biotinylated CCL5 Detector Antibody 120 uL 100X Biotinylated CCL5 Detector Antibody 120 uL Avidin/HRP Conjugate 120 uL Avidin/HRP Conjugate 120 uL Standard Diluent 1 x 20 mL Standard Diluent 1 x 20 mL Detector Antibody Diluent 1 x 12 mL Detector Antibody Diluent 1 x 12 mL Conjugate Diluent 1 x 12 mL Conjugate Diluent 1 x 12 mL 30X Wash Buffer 1 x 20 mL 30X Wash Buffer 1 x 20 mL TMB Substrate 1 x 9 mL TMB Substrate 1 x 9 mL Stop Solution 1 x 6 mL Stop Solution 1 x 6 mL

This assay has high sensitivity and excellent specificity for detection of Regulated On Activation In Normal T-Cell Expressed And Secreted (RANTES) . No significant cross-reactivity or interference between Regulated On Activation In Normal T-Cell Expressed And Secreted (RANTES) and analogues was observed.