应用说明
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.
Default 3
8
形式
Lyophilized powder
存储溶液
Reconstitute with 20mM Tris and 150mM NaCl to 0.1-1.0mg/ml. Do not vortex. Lyophilized from 20mM Tris, 150mM NaCl, 1mM EDTA, 1mM DTT, 0.01% SKL, 5% Trehalose.
保存剂
ProClin 300
存放说明
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)
表达系统
E. coli
纯度
> 97%
内毒素
< 1 EU/μg
偶联
Unconjugated
注意事项
For laboratory use only. Not for any clinical, therapeutic, or diagnostic use in humans or animals. Not for animal or human consumption.
别名
CD14 molecule
功能与背景
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]
数据库
研究领域