Human IL-8/CXCL8 ELISA Kit

Catalog Number: abs510004 Application: ELISA Reactivity: Human Conjugation:

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$450 USD

Size: 96T

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Product Details

Product Specification

Species Reactivity

Human

Theory

This kit adopts double antibody sandwich enzyme-linked immunosorbent detection technology. Specific anti-human IL-8 antibodies were pre-coated on high affinity plates. The standard substance, the sample to be tested and the biotinylated detection antibody are added to the well of the enzyme label plate, and after incubation, IL-8 present in the sample binds to the solid phase antibody and the detection antibody to form an immune complex. After washing to remove unbound material, horseradish peroxidase-labeled Streptavidin-HRP was added. After washing, a chromogenic substrate is added to protect the color from light. A stop solution was added to stop the reaction, and the absorbance value was measured at a wavelength of 450 nm (reference calibration wavelength of 540 nm or 570 nm).

Synonym

3-10C, AMCF-I, C-X-C motif chemokine 8, CXCL8, CXCL8SCYB8, Emoctakin, GCP1, GCP-1TSG-1, IL8, IL-8, interleukin 8, K60, LAI, LECT, MDNCF, MDNCFb-ENAP, member 8, MONAPGCP1, NAP1, NAP-1NAP1, NCF, Neutrophil-activating protein 1, Protein 3-10C, T cell chemotactic factor, T-cell chemotactic factor, TCF, TSG1

Composition

Please use it within the validity period of the kit (new and old products are shipped randomly)

form	Specifications	Shelf life of diluted or redissolved reagent after unpacking
Human IL-8/CXCL8 Microplate	1 piece	Unused slats can be stored at 2-8 °C for 30 days after being sealed back in aluminum foil bags with desiccant
Human IL-8/CXCL8 Standard	2 sticks	After dissolution, the calculated amount is dispensed and stored at-20 °C for 14 days
Human IL-8/CXCL8 detection antibody	1 stick	After dissolution in concentrated volume, it can be stored at 2-8 °C for 14 days
40 × SA-HRP	1 stick	40 × concentration can be stored at 2-8 ° C.; 1 × working concentration Not recommended for storage
10 × Buffer for concentration and dilution	1 vial	After opening, it can be stored at 2-8 °C for 30 days
Chromogenic liquid	1 vial
Stop liquid	1 vial
20 × concentrated wash buffer	1 vial
Sealing film	3 sheets	Stored at room temperature, to avoid contamination, not reusable

Background

Interleukin-8 (IL-8, also known as GCP-1, NAP-1, and CXCL8) is an 8-9kDa chemokine belonging to the α-type or CXC family that binds to heparin. So far, a total of 15 human CXC family proteins have been found, ranging in size from 8 to 12kDa. Most of them are located on human chromosome 4, all have a typical triple β-sheet layer/an α-helix structure, and most of them show Glu-Leu-Arg tripeptide motif at the N-terminus. Human IL-8 was first synthesized as a 99 amino acid precursor containing a 20 amino acid signal sequence and a 79 amino acid mature region. It can be circulated in vivo as monomers, homodimers, and heterodimers formed with CXCL4/PF4. IL-8 monomers are considered to be the most biologically active, whereas heterodimers may enhance the activity of PF4. At the amino acid level, mature human IL-8 has 65% and 70% homology to pigs and dogs, respectively. In rodents, the corresponding gene for IL-8 has not been found. A variety of IL-8 isoforms were generated by alternative splicing and differential proteolysis. Alternative splicing occurs at the C-terminus, where there is an 11 amino acid substitution (at aa # 92-99). Protease hydrolysis may be a cell-specific event that produces a truncation at the N-terminus of IL-8. For example, fibroblasts and vascular endothelial cells cleave amino acids 21 and 22 to form IL-8, while monocytes and lymphocytes cleave amino acids 21-25 to produce IL-8. These short IL-8s generally have higher biological activity, especially against the CXCR1IL-8 receptor. Approximately 15% of IL-8 is citrullinated at the precursor Arg27 position, which increases the half-life of IL-8 and promotes leukocytosis. Many types of cells secrete IL-8, including monocytes and neutrophils, fibroblasts and keratinocytes, mast cells, visceral smooth muscle cells, dendritic cells, type II large alveolar cells, and endothelial cells.
There are two IL-8 receptors, both of which belong to G protein-coupled receptor proteins: CXCR1/IL-8RA and CXCR2/IL-8RB, and the amino acid sequences between them share 77% homology. CXCR1 has a molecular weight of 45-50 kDa and is almost entirely exclusive to IL-8; CXCR2 has a molecular weight of 35-40 kDa and is common to all CXC chemokines. CXCR1 and CXCR2, respectively, form constitutive homodimers, which appear to be their functional configuration. Heterodimers also form when expressed in the same cell; But when it binds to IL-8, it will be disintegrated. CXCR2 responds to low concentrations of IL-8 and is mainly related to chemotaxis and MMP-9 release. In contrast, CXCR1 responds to high concentrations of IL-8 and is associated with respiratory bursts and activation of phospholipase D2. Therefore, in neutrophils CXCR2 is thought to direct the migration of neutrophils to the site of inflammation and then elicit CXCR1-mediated antibacterial activity.
The best known function of IL-8 is its pro-inflammatory effect in immune cells. Essentially, IL-8 is secreted by a variety of cell types exposed to inflammatory stimulatory factors. For monocytes/macrophages, microbial exposure causes the release of IL-8; Subsequently, CXCR2-mediated chemotaxis migrates neutrophils to the site of antigen attack, accompanied by the activation and initiation of antibacterial activity in the next step. IL-8 can enhance the effect of M-CSF in bone marrow, thereby causing the maturation and release of granulocytes. These two functions of IL-8 complement each other. It has been reported that IL-8 has angiogenic function on tumor-associated endothelial cells. At this time, tumor-derived IL-8 can activate CXCR1 and CXCR2 in vascular endothelial cells in a paracrine manner. Both CXCR1 and CXCR2 are associated with the PI3-K/Akt and RasGTP signaling pathways and are involved in cell survival and proliferation. In addition, IL-8 positively regulates VEGFR2 and EGFR, which belong to receptor tyrosine kinases that mediate cell growth and migration.

General Notes

1. Please use the kit within the validity period.
2. The components of different kits and different batch kits cannot be mixed.
3. If the sample value is greater than the highest value of the standard curve, the sample should be diluted with diluent (1 ×) and re-tested; If the cell culture supernatant sample needs to be distributed and diluted, cell culture medium can be used for other intermediate dilutions except dilution with diluent in the last step.
4. Differences in test results can be caused by a variety of factors, including the operation of the experimenter, the use of the pipette, the plate washing technique, the reaction time or temperature, the storage of the kit, etc.
5. The terminating solution in the kit is an acidic solution. Please protect your glasses, hands, face and clothes when using it.
6. For scientific research only, not for in vitro diagnosis.

Storage Temp.

Kit unopened, stored at 2-8 ℃.

Test Range

31.3pg/mL-2000pg/mL

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Human IL-8/CXCL8 ELISA Kit