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Human CXCL8 / IL8 protein (active)

Human CXCL8 / IL8 protein (active). GTX00228-pro
Human CXCL8 / IL8 protein (active). GTX00228-pro
Human CXCL8 / IL8 protein (active). GTX00228-pro
Human CXCL8 / IL8 protein (active). GTX00228-pro

Cat No. GTX00228-pro

Application Functional Assay
Species Human
Package
10 μg ($239)
APPLICATION

Application Note

Interleukin 8 (IL8 or chemokine (C-X-C motif) ligand 8, CXCL8) is a chemokine produced by macrophages and other cell types such as epithelial cells, airway smooth muscle cellsendothelial cells.   IL-8, also known as neutrophil chemotactic factor, has two primary functions. It induces chemotaxis in target cells, primarily neutrophils but also other granulocytes, causing them to migrate toward the site of infection. IL8 also induces phagocytosis once they have arrived. IL8 is also known to be a potent promoter of angiogenesis. Besides, Syndecan 1 (SDC1) has been identified as an interactor of IL8, thus a binding ELISA assay was conducted to detect the interaction of recombinant human IL8 and recombinant human SDC1. Briefly, IL8 were diluted serially in PBS, with 0. 01% BSA (pH 7. 4). Duplicate samples of 100 μl were then transferred to SDC1-coated microtiter wells and incubated for 2h at 37ºC. Wells were washed with PBST and incubated for 1h with anti-IL8 pAb, then aspirated and washed 3 times. After incubation with HRP labelled secondary antibody, wells were aspirated and washed 3 times. With the addition of substrate solution, wells were incubated 15-25 minutes at 37ºC. Finally, add 50 μl stop solution to the wells and read at 450nm immediately. The binding activity of IL8 and SDC1 was in a dose dependent manner.
IL8 is a kind of neutrophil chemotactic factor, so chemotaxis  assay used 24-well microchemotaxis system was undertaken to detect the chemotactic effect of IL8 on the human T-lymphocyte leukemia cell line Jurkat. Briefly, Jurkat cells were seeded into the upper chambers (100 μl cell suspension, 1x10⁶cells/ml in RPMI-1640 with FBS free) and recombinant human IL8 (10 ng/ml, 100 ng/ml and 1000 ng/ml diluted separately in serum free RPMI-1640) was added in lower chamber with a polycarbonate filter (8μm pore size) used to separate the two compartments. After incubation at 37ºC with 5% CO₂ for 1h, the filter was removed, then cells in low chamber were observed by inverted microscope at low magnification (×100) and the number of migrated cells were counted at high magnification (×400) randomly. The migrated Jurkat cells in low chamber at low magnification (×100) randomly, and the migrated cells were counted at high magnification (×400). IL8 is able to induce migration of Jurkat cells, and the optimum chemotaxis of IL8 occurs at 10~100 ng/ml.

Observed MW

12 kDa.
PROPERTIES

Form

Lyophilized powder

Buffer

Lyophilized from 20 mM Tris (pH 8.0) with 150 mM NaCl, 1 mM EDTA, 1 mM DTT, 0.01% SKL, 5% Trehalose, Proclin300. Reconstitute with 20 mM Tris and 150 mM NaCl (pH 8.0) to a concentration of 0.1-1.0 mg/mL. Do not vortex.

Storage

For short-term storage (1-2 weeks), store at 4ºC. For long-term storage, store at -20ºC or below. After reconstitution, keep as concentrated solution.Avoid freeze-thaw cycles.

Region/Sequence

N-terminal His-Tag; Ser28~Ser99 (NP_000575.1)

Expression System

E. coli

Purity

> 95%

Endotoxin

< 1 EU/μg

Conjugation

Unconjugated

Note

For laboratory use only. Not for any clinical, therapeutic, or diagnostic use in humans or animals. Not for animal or human consumption.
TARGET

Synonyms

C-X-C motif chemokine ligand 8,GCP-1,GCP1,IL8,LECT,LUCT,LYNAP,MDNCF,MONAP,NAF,NAP-1,NAP1

Background

The protein encoded by this gene is a member of the CXC chemokine family and is a major mediator of the inflammatory response. The encoded protein is secreted primarily by neutrophils, where it serves as a chemotactic factor by guiding the neutrophils to the site of infection. This chemokine 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 members of the CXC chemokine gene family form a gene cluster in a region of chromosome 4q. [provided by RefSeq, Aug 2017]

Database

Research Area

DATA IMAGES
Human CXCL8 / IL8 protein (active). GTX00228-pro

GTX00228-pro Functional Assay Image

The chemotactic effect of GTX00228-pro Human CXCL8 / IL8 protein (active).
Upper: (A) Jurkat cells were seeded into the upper chambers and serum free RPMI-1640 without IL8 was added in lower chamber, then cells in lower chamber were observed at low magnification (×100) after incubation for 1hr.
(B) Jurkat cells were seeded into the upper chambers and serum free RPMI-1640 with 10 ng/ml IL8 was added in lower chamber, then cells in lower chamber were observed at low magnification (×100) after incubation for 1hr.
Lower: The number of migrated cells were counted at high magnification (×400) randomly (five fields for each filter).

Human CXCL8 / IL8 protein (active). GTX00228-pro

GTX00228-pro Functional Assay Image

Functional ELISA analysis of GTX00228-pro Human CXCL8 / IL8 protein (active) which can bind immobilized SDC1 protein.

Human CXCL8 / IL8 protein (active). GTX00228-pro

GTX00228-pro Image

SDS-PAGE analysis of GTX00228-pro Human CXCL8 / IL8 protein (active).

Human CXCL8 / IL8 protein (active). GTX00228-pro

GTX00228-pro Image

WB analysis of GTX00228-pro Human CXCL8 / IL8 protein (active).

REFERENCE
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REVIEW
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