Journal: The Journal of biological chemistry

Article Title: Inhibition of macrophage scavenger receptor activity by tumor necrosis factor-alpha is transcriptionally and post-transcriptionally regulated.

doi: 10.1074/jbc.271.13.7767

Figure Lengend Snippet: FIG. 4. TNF-a decreases MSR mRNA synthesis in PMA-differ- entiated THP-1 macrophages at steady state level. A, total RNA was isolated from THP-1 cells grown in serum-free medium (Control, THP-1 monocytes, samples 1 and 5), medium containing PMA (150 nM) (PMA, THP-1 macrophages, samples 2 and 6), medium containing PMA (150 nM) 1 TNF-a (200 units/ml) (PMA 1 TNF-a, samples 3 and 7), or medium containing TNF-a (200 units/ml) (TNF-a, samples 4 and 8) for 20 and 48 h, respectively. Northern blots were hybridized with 32P- labeled cDNAs of MSR or 28 S. The respective autoradiograms with MSR and 28 S bands are labeled and indicated with arrows. B, histo- grams represent quantification by PhosphorImager® of MSR mRNA normalized by comparison with 28 S ribosomal RNA. All data are expressed as a percentage of sample 2, i.e. PMA-differentiated THP-1 macrophages. Similar results were obtained in three separate experi- ments. C, RT-PCR analysis of the expression of macrophage scavenger receptor types SR-AI and SR-AII mRNA in THP-1 monocytes, PMA- differentiated THP-1 macrophages in the absence and the presence of TNF-a. Ethidium bromide-stained agarose gel with MSR type SR-AI at 447 base pairs and MSR type SR-AII at about 285 base pairs. Lane 1, 100 base pair DNA ladder; lane 2, THP-1 monocytes; lane 3, PMA- differentiated THP-1 macrophages (24 h); lane 4, THP-1 cells with PMA 1 TNF-a (24 h); lane 5, DNA molecular weight marker V from Boeh- ringer Mannheim. The detailed method is described under ‘‘Experimen- tal Procedures.’’ There was no detectable MSR type SR-AI and SR-AII mRNA in THP-1 cells with TNF-a (data not shown). The arrows indi- cate MSR types SR-AI and SR-AII. The results shown are representa- tive of three independent experiments.

Article Snippet: Medium RPMI 1640 medium, L-glutamine, penicillin, streptomycin, and fetal calf serum were purchased from Life Technologies, Inc. A 100-base pair DNA ladder was purchased from Life Technologies, Inc., MD.

Techniques: Isolation, Control, Northern Blot, Labeling, Comparison, Reverse Transcription Polymerase Chain Reaction, Expressing, Staining, Agarose Gel Electrophoresis, Molecular Weight, Marker

Journal: Biosensors

Article Title: Ultrasensitive Label-Free Sensing of IL-6 Based on PASE Functionalized Carbon Nanotube Micro-Arrays with RNA-Aptamers as Molecular Recognition Elements

doi: 10.3390/bios7020017

Figure Lengend Snippet: Schematic showing the reaction of 1-Pyrenebutanoic Acid Succinimidyl Ester (PASE) onto IL-6 aptamer forming the IL-6 aptamer PASE complex. Three groups of devices were prepared, Carbon Nanotube (CNT) device functionalized with ( B ) IL-6 aptamer PASE complex, positive control; ( C ) IgG, negative control; and ( D ) tween-20 blocking agent, negative control.

Article Snippet: CNT biosensors were functionalized with 1-Pyrenebutanoic Acid Succinimidyl Ester (PASE) conjugated IL-6 aptamers (Base Pair biotechnologies, Pearland, TX, USA; Cat.#: ATW0077) at 20 μg/mL in 1 mM MgCL 2 1× PBS buffer solution for 2 h at room temperature.

Techniques: Positive Control, Negative Control, Blocking Assay

Journal: Biosensors

Article Title: Ultrasensitive Label-Free Sensing of IL-6 Based on PASE Functionalized Carbon Nanotube Micro-Arrays with RNA-Aptamers as Molecular Recognition Elements

doi: 10.3390/bios7020017

Figure Lengend Snippet: Atomic Force Microscope (AFM) image showing a phase image ( A ); and Z -Axis image ( B ), of bare CNT ( a , b ), functionalized CNT with IL-6 aptamer ( c , d ), and IL-6 protein binding to aptamer/CNT structure ( e , f ). Images were obtained using dry tapping mode (Nanosurf NaioAFM system). Scale bar = 100 nm.

Article Snippet: CNT biosensors were functionalized with 1-Pyrenebutanoic Acid Succinimidyl Ester (PASE) conjugated IL-6 aptamers (Base Pair biotechnologies, Pearland, TX, USA; Cat.#: ATW0077) at 20 μg/mL in 1 mM MgCL 2 1× PBS buffer solution for 2 h at room temperature.

Techniques: Microscopy, Protein Binding

Journal: Biosensors

Article Title: Ultrasensitive Label-Free Sensing of IL-6 Based on PASE Functionalized Carbon Nanotube Micro-Arrays with RNA-Aptamers as Molecular Recognition Elements

doi: 10.3390/bios7020017

Figure Lengend Snippet: Sensor response of the CNT-biosensor at source-drain bias of 0.1 V and at the gate bias of 0 V. ( A ) indicates the response after the introduction of 20 nM BSA (red line), 1 mM MgCL 2 1× PBS (gray line), 1 pg IL-6 target protein (blue line) to functionalized CNT-biosensor and ( B ) 1 pg IL-6 target protein to bare unfunctionalized CNT-biosensor (green line); ( C ) Response of tween-20 blocked CNT-biosensor to IL-6 protein (blue line), and BSA (red line). Arrow indicates the point of sample injection.

Article Snippet: CNT biosensors were functionalized with 1-Pyrenebutanoic Acid Succinimidyl Ester (PASE) conjugated IL-6 aptamers (Base Pair biotechnologies, Pearland, TX, USA; Cat.#: ATW0077) at 20 μg/mL in 1 mM MgCL 2 1× PBS buffer solution for 2 h at room temperature.

Techniques: Injection

Journal: Biosensors

Article Title: Ultrasensitive Label-Free Sensing of IL-6 Based on PASE Functionalized Carbon Nanotube Micro-Arrays with RNA-Aptamers as Molecular Recognition Elements

doi: 10.3390/bios7020017

Figure Lengend Snippet: Before and after I DS -V GS characterization of IL-6 aptamer functionalized CNT-biosensor with ( A ) blank 1 mM MgCL 2 1× PBS samples; ( B ) 20 nM BSA; and ( C ) 1 pg and 1 ng IL-6 protein injection. I DS -V GS response of tween-20 blocked CNT-biosensor to BSA ( D ); and IL-6 protein ( E ).

Article Snippet: CNT biosensors were functionalized with 1-Pyrenebutanoic Acid Succinimidyl Ester (PASE) conjugated IL-6 aptamers (Base Pair biotechnologies, Pearland, TX, USA; Cat.#: ATW0077) at 20 μg/mL in 1 mM MgCL 2 1× PBS buffer solution for 2 h at room temperature.

Techniques: Injection

Journal: Biosensors

Article Title: Ultrasensitive Label-Free Sensing of IL-6 Based on PASE Functionalized Carbon Nanotube Micro-Arrays with RNA-Aptamers as Molecular Recognition Elements

doi: 10.3390/bios7020017

Figure Lengend Snippet: ( A ) Time dependence of normalized source-drain current of the CNT-biosensor at the source-drain bias of 0.1 V and at gate bias of 0 V after the introduction of IL-6 target protein at various concentrations onto the IL-6 aptamer functionalized CNT-biosensor. Arrows indicate the points of IL-6 target protein injection; ( B ) The Sensitivity of IL-6 aptamer CNT-biosensor as a function of IL-6 protein concentration (1 pg to 10 ng /mL). The inner box demonstrates the data in logarithmic scale.

Article Snippet: CNT biosensors were functionalized with 1-Pyrenebutanoic Acid Succinimidyl Ester (PASE) conjugated IL-6 aptamers (Base Pair biotechnologies, Pearland, TX, USA; Cat.#: ATW0077) at 20 μg/mL in 1 mM MgCL 2 1× PBS buffer solution for 2 h at room temperature.

Techniques: Injection, Protein Concentration

Journal: Biosensors

Article Title: Ultrasensitive Label-Free Sensing of IL-6 Based on PASE Functionalized Carbon Nanotube Micro-Arrays with RNA-Aptamers as Molecular Recognition Elements

doi: 10.3390/bios7020017

Figure Lengend Snippet: Blood experiment results. Response of the CNT-biosensor at source-drain bias of 0.1 V and at the gate bias of 0 V. IL-6 spiked blood samples were tested on two different CNT-biosensor surfaces, IL-6 aptamer ( A ), and IgG ( B ) functionalized surfaces. The response of the IL-6 aptamer functionalized devices to control buffer, 1 mM MgCl 2 1× PBS, is presented in each panel as a reference point (black marks). ( A ) Shows the respond of the IL-6 aptamer functionalized device to IL-6 protein spiked blood (10 pg/mL), ~8–13% drop in the current (diagnostic zone); ( B ) Presents the response to IgG functionalized CNT-biosensor, negative control surface, to IL-6 spiked blood showing no significant change in signal. Arrows indicate the points of IL-6 spiked blood injection, at 60 s.

Article Snippet: CNT biosensors were functionalized with 1-Pyrenebutanoic Acid Succinimidyl Ester (PASE) conjugated IL-6 aptamers (Base Pair biotechnologies, Pearland, TX, USA; Cat.#: ATW0077) at 20 μg/mL in 1 mM MgCL 2 1× PBS buffer solution for 2 h at room temperature.

Techniques: Diagnostic Assay, Negative Control, Injection

Journal: Biosensors

Article Title: Ultrasensitive Label-Free Sensing of IL-6 Based on PASE Functionalized Carbon Nanotube Micro-Arrays with RNA-Aptamers as Molecular Recognition Elements

doi: 10.3390/bios7020017

Figure Lengend Snippet: ( A ) Time dependence of normalized source-drain current of the CNT-biosensor at the source-drain bias of 0.1 V and at gate bias of 0 V after the introduction of IL-6 spiked blood sample at various concentrations onto the IL-6 aptamer functionalized CNT-biosensor. Arrows indicate the points of IL-6 spiked blood injection; ( B ) The Sensitivity of IL-6 aptamer CNT-biosensor as a function of IL-6 protein concentration in blood sample (10 pg to 1 ng /mL) in logarithmic scale. Diagnostic gray zone for large tumor mass at ~12–40 pg/mL is highlighted in yellow which is higher than our sensor’s lower limit sensing threshold at 10 pg/mL.

Article Snippet: CNT biosensors were functionalized with 1-Pyrenebutanoic Acid Succinimidyl Ester (PASE) conjugated IL-6 aptamers (Base Pair biotechnologies, Pearland, TX, USA; Cat.#: ATW0077) at 20 μg/mL in 1 mM MgCL 2 1× PBS buffer solution for 2 h at room temperature.

Techniques: Injection, Protein Concentration, Diagnostic Assay