Active Lipopolysaccharide Binding Protein (LBP)

LPS-Binding Protein

ACTIVITY TEST

Figure. The binding activity of LBP with CD14.
Lipopolysaccharide Binding Protein (LBP) is a soluble acute-phase protein that binds to bacterial lipopolysaccharide (or LPS) to elicit immune responses by presenting the LPS to important cell surface pattern recognition receptors called CD14 and TLR4. The protein encoded by this gene is involved in the acute-phase immunologic response to gram-negative bacterial infections. This protein is part of a family of structurally and functionally related proteins, including BPI, plasma cholesteryl ester transfer protein (CETP), and phospholipid transfer protein (PLTP). Besides, Cluster Of Differentiation 14 (CD14) has been identified as an interactor of LBP, thus a binding ELISA assay was conducted to detect the interaction of recombinant human LBP and recombinant human CD14. Briefly, LBP were diluted serially in PBS, with 0.01% BSA (pH 7.4). Duplicate samples of 100µL were then transferred to CD14-coated microtiter wells and incubated for 2h at 37℃. Wells were washed with PBST and incubated for 1h with anti-LBP 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 37℃. Finally, add 50µL stop solution to the wells and read at 450nm immediately. The binding activity of LBP and CD14 was shown in Figure 1, and this effect was in a dose dependent manner.

USAGE

Reconstitute in 20mM Tris, 150mM NaCl (pH8.0) to a concentration of 0.1-1.0 mg/mL. Do not vortex.

STORAGE

Avoid repeated freeze/thaw cycles. Store at 2-8°C for one month. Aliquot and store at -80°C for 12 months.

STABILITY

The thermal stability is described by the loss rate. The loss rate was determined by accelerated thermal degradation test, that is, incubate the protein at 37°C for 48h, and no obvious degradation and precipitation were observed. The loss rate is less than 5% within the expiration date under appropriate storage condition.

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Magazine Citations
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