Journal: Soft matter

Article Title: Membrane curvature recognition by C-reactive protein using lipoprotein mimics

doi: 10.1039/C2SM25779C

Figure Lengend Snippet: Binding of CRP-LPP mimics to immobilized C1q by ELISA. Purified human C1q (1 μg/mL) was coated on ELISA plates and the plates were incubated at increasing concentrations with each CRP-LPP mimic: 21 nm ( ), 26 nm ( ), 28 nm ( ), 41 nm ( ), 64 nm ( ), and PC liposome (□). The degree of mCRP binding was detected using biotinylated polyclonal anti-CRP antibody. The measured absorbance was normalized to a BSA blank control. (a) Concentration-dependent binding of CRP-LPP mimic to C1q. Inset: C1q binding of the pCRP (×) and mCRP (○) controls. (b) Binding of CRP-LPP mimics (1.8 μg/mL CRP) to C1q. Data represents mean absorbance ±SD, n = 3. ** p<0.01; ***p<0.001 compared to pCRP.

Article Snippet: Purified human C1q (1 μg/mL) was diluted in coating buffer (30 mM Na 2 CO 3 , 70 mM NaHCO 3 , pH 9.6, Sigma-Aldrich) and incubated overnight at 4°C on high binding Microlon® ELISA plates (Santa Cruz Biotechnology).

Techniques: Binding Assay, Enzyme-linked Immunosorbent Assay, Purification, Incubation, Control, Concentration Assay

Journal: Soft matter

Article Title: Membrane curvature recognition by C-reactive protein using lipoprotein mimics

doi: 10.1039/C2SM25779C

Figure Lengend Snippet: Competitive binding of CRP to oxLDL using LPP mimics. Air oxidized LDLs (5.7 μg/mL) were immobilized on ELISA plates overnight at 4°C. Equal amounts of CRP (3.6 μg/mL) were pre-incubated for 30 min with LPP mimics: 21 nm, 26 nm, 28 nm, 41 nm, 64 nm, and PC liposome, or PCh-BSA, pCRP and mCRP controls. Then the CRP-LPP mimics were added to the oxLDL-coated plates. The binding of CRP to oxLDL was detected using biotinylated polyclonal anti-CRP antibody and the measured absorbance was normalized to pCRP. Inset: Competitive immunoassay using PCh-BSA. Data represents mean absorbance ±SD, n = 5. * p<0.05; ***p<0.001 compared to pCRP.

Article Snippet: Purified human C1q (1 μg/mL) was diluted in coating buffer (30 mM Na 2 CO 3 , 70 mM NaHCO 3 , pH 9.6, Sigma-Aldrich) and incubated overnight at 4°C on high binding Microlon® ELISA plates (Santa Cruz Biotechnology).

Techniques: Binding Assay, Enzyme-linked Immunosorbent Assay, Incubation

Journal: Soft matter

Article Title: Membrane curvature recognition by C-reactive protein using lipoprotein mimics

doi: 10.1039/C2SM25779C

Figure Lengend Snippet: Representative topographic AFM images of LDL immobilized on ELISA well. AFM images of (A) high binding ELISA well, (B) antibody-captured LDLs and (C) direct immobilized LDLs on ELISA well. LDLs (5.7 μg/mL) were captured using a anti-Apo B polyclonal antibody or directly immobilized onto high binding ELISA well and incubated overnight at 4°C. After rinsing and drying, images were acquired in air using a tapping mode AFM. Insets show a cartoon illustrating the probable structures of the respective surfaces. Image scan size = 5 × 5μm.

Article Snippet: Purified human C1q (1 μg/mL) was diluted in coating buffer (30 mM Na 2 CO 3 , 70 mM NaHCO 3 , pH 9.6, Sigma-Aldrich) and incubated overnight at 4°C on high binding Microlon® ELISA plates (Santa Cruz Biotechnology).

Techniques: Enzyme-linked Immunosorbent Assay, Binding Assay, Incubation