Human IL8 CLIA Kit (HUES00047)
Human IL8 (Interleukin 8) CLIA Kit
This kit uses Sandwich-CLIA as the method. The micro CLIA plate provided in this kit has been pre-coated with an antibody specific to Human IL8 . Standards or samples are added to the appropriate micro CLIA plate wells and combined with the specific antibody. Then a biotinylated detection antibody specific for Human IL8 and Avidin-Horseradish Peroxidase (HRP) conjugate are added to each micro plate well successively and incubated. Free components are washed away. The substrate solution is added to each well. Only those wells that contain Human IL8, biotinylated detection antibody and Avidin-HRP conjugate will appear fluorescence. The Relative light unit (RLU) value is measured spectrophotometrically by the Chemiluminescence immunoassay analyzer. The RLU value is positively associated with the concentration of Human IL8. The concentration of Human IL8 in the samples can be calculated by comparing the RLU of the samples to the standard curve.
|Detection range||15.63-1000 pg/mL|
|Sample type||Serum, plasma and other biological fluids|
|Repeatability||CV < 15%|
This kit recognizes Human IL8 CLIA Kit in samples. No significant cross-reactivity or interference between Human IL8 CLIA Kit and analogues was observed.
As the RLU values of the standard curve may vary according to the conditions of the actual assay performance (e.g. operator, pipetting technique, washing technique or temperature effects), the operator should establish a standard curve for each test. Typical standard curve and data is provided below for reference only.
Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and high level Human IL8 CLIA Kit were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level Human IL8 CLIA Kit were tested on 3 different plates, 20 replicates in each plate.
|Intra-assay Precision||Inter-assay Precision|
|C V (%)||9.82||10.77||6.96||11.71||8.01||6.77|
The recovery of Human IL8 CLIA Kit spiked at three different levels in samples throughout the range of the assay was evaluated in various matrices.
|Sample Type||Range (%)||Average Recovery (%)|
|EDTA plasma (n=5)||100-115||107|
|Cell culture media (n=5)||99-115||106|
Samples were spiked with high concentrations of Human IL8 CLIA Kit and diluted with Reference Standard & Sample Diluent to produce samples with values within the range of the assay.
|Serum (n=5)||EDTA plasma (n=5)||Cell culture media (n=5)|
Kit Components & Storage
An unopened kit can be stored at 4'C for 1 month. If the kit is not used within 1 month, store the items separately according to the following conditions once the kit is received.
|Micro CLIA Plate(Dismountable)||8 wells X12 strips||-20'C, 6 months|
|Reference Standard||2 vials|
|Concentrated Biotinylated Detection Ab (100X)||1 vial, 120 uL|
|Concentrated HRP Conjugate (100X)||1 vial, 120 uL||-20'C(shading light), 6 months|
|Reference Standard & Sample Diluent||1 vial, 20 mL||4'C, 6 months|
|Biotinylated Detection Ab Diluent||1 vial, 14 mL|
|HRP Conjugate Diluent||1 vial, 14 mL|
|Concentrated Wash Buffer (25X)||1 vial, 30 mL|
|Substrate Reagent A||1 vial, 5 mL||4'C (shading light)|
|Substrate Reagent B||1 vial, 5 mL||4'C (shading light)|
|Plate Sealer||5 pieces|
|Product Description||1 copy|
|Certificate of Analysis||1 copy|
Human IL8 CLIA Kit (HUES00047) Assay procedure
- 1. Set standard, test sample and control (zero) wells on the pre-coated plate and record theirpositions. It is recommended to measure each standard and sample in duplicate. Note: addall solutions to the bottom of the plate wells while avoiding contact with the well walls. Ensuresolutions do not foam when adding to the wells.
- 2. Aliquot 100µl of standard solutions into the standard wells.
- 3. Add 100µl of Sample / Standard dilution buffer into the control (zero) well.
- 4. Add 100µl of properly diluted sample (serum, plasma, tissue homogenates and otherbiological fluids.) into test sample wells.
- 5. Cover the plate with the sealer provided in the kit and incubate for 90 min at 37°C.
- 6. Aspirate the liquid from each well, do not wash. Immediately add 100µL of BiotinylatedDetection Ab working solution to each well. Cover the plate with a plate seal and gently mix.Incubate for 1 hour at 37°C.
- 7. Aspirate or decant the solution from the plate and add 350µL of wash buffer to each welland incubate for 1-2 minutes at room temperature. Aspirate the solution from each well andclap the plate on absorbent filter paper to dry. Repeat this process 3 times. Note: a microplatewasher can be used in this step and other wash steps.
- 8. Add 100µL of HRP Conjugate working solution to each well. Cover with a plate seal andincubate for 30 min at 37°C.
- 9. Aspirate or decant the solution from each well. Repeat the wash process for five times asconducted in step 7.
- 10. Add 100µL of Substrate mixture solution to each well. Cover with a new plate seal andincubate for no more than 5 min at 37°C. Protect the plate from light.
- 11. Determine the RLU value of each well immediately.
Human IL8 CLIA Kit (HUES00047) Protein Information
|UniProt Protein Function:||IL8: IL-8 is a chemotactic factor that attracts neutrophils, basophils, and T-cells, but not monocytes. It is also involved in neutrophil activation. It is released from several cell types in response to an inflammatory stimulus. IL-8(6-77) has a 5-10-fold higher activity on neutrophil activation, IL-8(5-77) has increased activity on neutrophil activation and IL-8(7-77) has a higher affinity to receptors CXCR1 and CXCR2 as compared to IL-8(1-77), respectively. Belongs to the intercrine alpha (chemokine CxC) family. 2 isoforms of the human protein are produced by alternative splicing.|
|UniProt Protein Details:|
Protein type:Secreted, signal peptide; Cytokine; Secreted; Motility/polarity/chemotaxis
Chromosomal Location of Human Ortholog: 4q13-q21
Cellular Component: extracellular space; extracellular region
Molecular Function:protein binding; chemokine activity; interleukin-8 receptor binding
Biological Process: regulation of cell adhesion; neutrophil chemotaxis; neutrophil activation; negative regulation of G-protein coupled receptor protein signaling pathway; unfolded protein response; calcium-mediated signaling; chemotaxis; signal transduction; regulation of retroviral genome replication; induction of positive chemotaxis; G-protein coupled receptor protein signaling pathway; negative regulation of cell proliferation; cellular protein metabolic process; unfolded protein response, activation of signaling protein activity; response to molecule of bacterial origin; receptor internalization; immune response; angiogenesis; cell cycle arrest; cell motility; inflammatory response; embryonic gut development
|NCBI Summary:||The protein encoded by this gene is a member of the CXC chemokine family. This chemokine is one of the major mediators of the inflammatory response. This chemokine is secreted by several cell types. It functions as a chemoattractant, and is also a potent angiogenic factor. This gene is believed to play a role in the pathogenesis of bronchiolitis, a common respiratory tract disease caused by viral infection. This gene and other ten members of the CXC chemokine gene family form a chemokine gene cluster in a region mapped to chromosome 4q. [provided by RefSeq, Jul 2008]|
|NCBI GenInfo Identifier:||124359|
|NCBI Gene ID:||3576|
|UniProt Secondary Accession:||P10145,Q6FGF6, Q6LAE6, Q96RG6, Q9C077, Q9UCE1, Q9UCR8 Q9UCR9, Q9UCS0, B2R4L8,|
|UniProt Related Accession:||P10145|
|Molecular Weight:||11,338 Da|
|NCBI Full Name:||Interleukin-8|
|NCBI Synonym Full Names:||chemokine (C-X-C motif) ligand 8|
|NCBI Official Symbol:||CXCL8|
|NCBI Official Synonym Symbols:||IL8; NAF; GCP1; LECT; LUCT; NAP1; GCP-1; LYNAP; MDNCF; MONAP; NAP-1|
|NCBI Protein Information:||interleukin-8; emoctakin; interleukin 8; T-cell chemotactic factor; neutrophil-activating peptide 1; beta-thromboglobulin-like protein; granulocyte chemotactic protein 1; tumor necrosis factor-induced gene 1; alveolar macrophage chemotactic factor I; monocyte-derived neutrophil chemotactic factor; monocyte-derived neutrophil-activating peptide; small inducible cytokine subfamily B, member 8; lymphocyte derived neutrophil activating peptide; lung giant cell carcinoma-derived chemotactic protein; beta endothelial cell-derived neutrophil activating peptide|
|UniProt Protein Name:||Interleukin-8|
|UniProt Synonym Protein Names:||C-X-C motif chemokine 8; Chemokine (C-X-C motif) ligand 8; Emoctakin; Granulocyte chemotactic protein 1; GCP-1; Monocyte-derived neutrophil chemotactic factor; MDNCF; Monocyte-derived neutrophil-activating peptide; MONAP; Neutrophil-activating protein 1; NAP-1; Protein 3-10C; T-cell chemotactic factorCleaved into the following 7 chains:MDNCF-aAlternative name(s):GCP/IL-8 protein IV; IL8/NAP1 form I|
|UniProt Gene Name:||CXCL8|
|UniProt Entry Name:||IL8_HUMAN|