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Human CD14 protein (active)

Human CD14 protein (active). GTX00249-pro
Human CD14 protein (active). GTX00249-pro
Human CD14 protein (active). GTX00249-pro
Human CD14 protein (active). GTX00249-pro

Cat No. GTX00249-pro

Application Functional Assay
Species Human
Package
10 μg ($279)
APPLICATION

Application Note

Cluster Of Differentiation 14 (CD14), also known as CD14, is a component of the innate immune system. CD14 acts as a co-receptor (along with the Toll-like receptor TLR 4 and MD-2) for the detection of bacterial lipopolysaccharide (LPS). CD14 can bind LPS only in the presence of lipopolysaccharide-binding protein (LBP). Although LPS is considered its main ligand, CD14 also recognizes other pathogen-associated molecular patterns such as lipoteichoic acid. Besides, Lipopolysaccharide Binding Protein (LBP) has been identified as an interactor of CD14, thus a binding ELISA assay was conducted to detect the interaction of recombinant human CD14 and recombinant human LBP. Briefly, CD14 were diluted serially in PBS with 0. 01% BSA (pH 7. 4). Duplicate samples of 100 μl were then transferred to LBP-coated microtiter wells and incubated for 2h at 37ºC. Wells were washed with PBST and incubated for 1h with anti-CD14 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 CD14 and LBP was in a dose dependent manner.
CD14 also can enhance LPS-stimulated IL-8 secretion by THP-1 human acute monocytic leukemia cells. Therefore, THP-1 cells were cultured in 24 well plates at a concentration of 1x10⁶ cells/ml and stimulated by LPS (1μg/ml), then added of different concentrations of recombinant human CD14 (1μg/ml, 5μg/ml). After 24h later, the production of IL8 was determined in the supernatants by cytokine specific ELISA. When the concentration of CD14 was 5μg/ml, the secretion LPS-stimulated IL-8 can be significantly increased.

Observed MW

18 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; Asp125~Leu288 (NP_000582.1)

Expression System

E. coli

Purity

> 97%

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

CD14 molecule

Background

The protein encoded by this gene is a surface antigen that is preferentially expressed on monocytes/macrophages. It cooperates with other proteins to mediate the innate immune response to bacterial lipopolysaccharide. Alternative splicing results in multiple transcript variants encoding the same protein. [provided by RefSeq, Mar 2010]

Database

Research Area

DATA IMAGES
Human CD14 protein (active). GTX00249-pro

GTX00249-pro Functional Assay Image

ELISA detection of secreted IL-8 protein activation from THP-1 cells treated with LPS (1μg/ml) and induced by GTX00249-pro Human CD14 protein (active) for 24hrs.
Recombinant protein concentrations: 5 μg/ml

Human CD14 protein (active). GTX00249-pro

GTX00249-pro Functional Assay Image

Functional ELISA analysis of GTX00249-pro Human CD14 protein (active) which can bind immobilized LBP protein.

Human CD14 protein (active). GTX00249-pro

GTX00249-pro Image

WB analysis of GTX00249-pro Human CD14 protein (active).

Human CD14 protein (active). GTX00249-pro

GTX00249-pro Image

SDS-PAGE analysis of GTX00249-pro Human CD14 protein (active).

REFERENCE
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Package List Price ($)
$ 279