Journal: Glycobiology

Article Title: Glycation does not modify bovine serum albumin (BSA)-induced reduction of rat aortic relaxation: The response to glycated and nonglycated BSA is lost in metabolic syndrome

doi: 10.1093/glycob/cwn034

Figure Lengend Snippet: Purification of AGE-BSA on Affi-Gel-Blue. Three peaks were observed: the first one corresponds to highly glycated BSA which did not bind to the matrix, the second one corresponds to the less glycated BSA which bound moderately, and the third peak corresponds to the stronger bound BSA. The first peak was the only one to be further analyzed. The arrows indicate the buffer used at each stage.

Article Snippet: Mass increase analysis of glycated BSA The mass change of BSA after glycation was analyzed by matrix-assisted laser desorption ionization mass spectrometry (MALDI-TOF) (Voyager DE-PRO from Applied Biosystems, CA), equipped with a nitrogen laser (337 nm), operating in a positive high-energy linear mode.

Techniques: Purification

Journal: Glycobiology

Article Title: Glycation does not modify bovine serum albumin (BSA)-induced reduction of rat aortic relaxation: The response to glycated and nonglycated BSA is lost in metabolic syndrome

doi: 10.1093/glycob/cwn034

Figure Lengend Snippet: Analysis of BSA and AGE-BSA by mass spectrometry. The mass increase of protein after glycation is observed. The mass of nonglycated BSA was 66,655.58 Da ( A ) and that of AGE-BSA was 74,461.15 Da ( B ). The net increase of mass was 7,805.57 Da, corresponding to the addition of 48 molecules of glucose to each BSA molecule. Inset: change of isoelectric point of BSA after glycation. Nonglycated BSA had a pI of 4.2 while AGE-BSA had a pI of 6.3. Line 2 is the broad range pI standard kit (pH 3–10).

Article Snippet: Mass increase analysis of glycated BSA The mass change of BSA after glycation was analyzed by matrix-assisted laser desorption ionization mass spectrometry (MALDI-TOF) (Voyager DE-PRO from Applied Biosystems, CA), equipped with a nitrogen laser (337 nm), operating in a positive high-energy linear mode.

Techniques: Mass Spectrometry

Journal: The Journal of Neuroscience

Article Title: Lewisx and α2,3-Sialyl Glycans and Their Receptors TAG-1, Contactin, and L1 Mediate CD24-Dependent Neurite Outgrowth

doi: 10.1523/JNEUROSCI.4361-08.2009

Figure Lengend Snippet: TAG-1 and Contactin bind to CD24 and are receptors for Lewis x . A , Lysates from CHO cells transfected with TAX-1 (CHO/TAX-1), Contactin (CHO/Contactin), or NCAM120 (CHO/NCAM120) were incubated with epoxy beads coated with Lewis x –BSA (Le x -BSA), N -acetyllactosamine–BSA (LN-BSA), and BSA. After pull-down, beads were analyzed by Western blotting using antibodies to TAG-1 (lanes 1–4), Contactin (lanes 5–8), or NCAM (lanes 9–12). B , Immunoprecipitates from brain homogenate using CD24 antibody (α-CD24) or control rat IgG (rIgG) were analyzed by Western blotting using antibodies against L1, TAG-1, Contactin, NCAM, or CD24. C , CD24-coupled epoxy beads were incubated with TAX-1-Fc, Contactin-Fc, CHL1-Fc, or Fc alone. After pull-down, beads (PD CD24) were analyzed by Western blotting using HRP-coupled anti-human antibody. D , E , Cerebellar tissue (input) was used for immunoprecipitations with CD24 with antibody (α-CD24), with control rat IgG (rIgG) or without antibody (−) or for immunoprecipitations with TAG-1 antibody (α-TAG-1), Contactin antibody (α-Contactin), or control goat IgG (gIgG) or without antibody (−). Immunoprecipitates were analyzed by Western blotting using antibodies against CD24, TAG-1, Contactin, or CHL1.

Article Snippet: For pull-down assays, 80 μg of Lewisx –BSA, N -acetyllactosamine–BSA, or BSA (Sigma) was coated onto Dynabeads M-270 Epoxy (Dynal) by overnight incubation at 4°C.

Techniques: Transfection, Incubation, Western Blot

Journal: The Journal of Biological Chemistry

Article Title: Apolipoprotein A-IV Reduces Hepatic Gluconeogenesis through Nuclear Receptor NR1D1 *

doi: 10.1074/jbc.M113.511766

Figure Lengend Snippet: ApoA-IV induces NR1D1 expression in primary mouse hepatocytes and HepG2 and HEK-293 cells. The cells were starved in DMEM with 1% BSA overnight and then treated with or without 20 μg/ml r-m-apoA-IV in mouse hepatocytes for the indicated times or 50 μg/ml r-h-apoA-IV in HepG2 and HEK-293 for 2 h. mRNA levels in primary mouse hepatocytes were quantitated by real-time RT-PCR and normalized to cyclophilin. NR1D1 protein levels were detected by Western immunoblotting with anti-NR1D1 antibody and then standardized against actin. The results are from three independent experiments with at least three samples each time. Data are presented as mean ± S.E. *, p

Article Snippet: Cells were seeded on 8-well chamber slides, maintained for 48 h with growth medium, serum-starved overnight in growth medium with 1% BSA, and then incubated with recombinant human apoA-IV protein fused with GFP (r-h-apoA-IV-GFP) or GFP only as a negative control for 2 h. After fixation, permeabilization, and blocking, the cells were incubated with anti-human NR1D1 (1:200) and anti-GFP (1:200) antibodies (Cell Signaling Technology, Beverly, MA) and then with Alexa Fluor 594-conjugated goat anti-rabbit and FITC-conjugated goat anti-mouse secondary antibodies (Invitrogen).

Techniques: Expressing, Quantitative RT-PCR, Western Blot

Journal: Scientific Reports

Article Title: Distinct Differentiation Programs Triggered by IL-6 and LPS in Teleost IgM+ B Cells in The Absence of Germinal Centers

doi: 10.1038/srep30004

Figure Lengend Snippet: IL-6 has no effect on the phagocytic capacity of IgM + B cells. Splenocytes were cultured in the presence of IL-6 (200 ng/ml) or LPS (100 μg/ml) for 24, 48 or 72 h at 20 °C. Non-stimulated controls were also included. After the different incubation periods, cells were exposed to fluorescent beads for a further 3 h at 20 °C. Non-ingested beads were removed by centrifugation over a cushion of 3% (w/v) BSA in PBS supplemented with 4.5 (w/v) D-glucose (Sigma). ( a ) Data are shown as mean percentage of phagocytic IgM + B cells (left) or Mean fluorescence intensity (MFI) (right) + standard deviation from six independent fish. * P

Article Snippet: Non-ingested beads were removed by centrifugation (100 × g for 10 min at 4 °C) over a cushion of 3% (w/v) BSA (Fraction V; Fisher Scientific) in PBS supplemented with 4.5 (w/v) D-glucose (Sigma).

Techniques: Cell Culture, Incubation, Centrifugation, Fluorescence, Standard Deviation, Fluorescence In Situ Hybridization

Journal: Medical Science Monitor : International Medical Journal of Experimental and Clinical Research

Article Title: Development and In Vitro Release of Isoniazid and Rifampicin-Loaded Bovine Serum Albumin Nanoparticles

doi: 10.12659/MSM.905581

Figure Lengend Snippet: In vitro release of isoniazid (INH: ) and rifampicin (RFP: ) from INH and RFP-loaded bovine serum albumin nanoparticles (INH-RFP-BSA-NPs) into culture medium.

Article Snippet: Preparation of INH-RFP-BSA-NPs An aqueous phase was prepared by thoroughly dissolving 0.4 g of INH (Wuhan Fuchi Biotechnology Co. Ltd., Wuhan, China) and 1.0 g of BSA (Amresco, Solon, OH, USA) in 200 ml of purified water at room temperature using ultrasound.

Techniques: In Vitro

Journal: Scientific Reports

Article Title: A rapid method for detecting protein-nucleic acid interactions by protein induced fluorescence enhancement

doi: 10.1038/srep39653

Figure Lengend Snippet: Detection of BamHI/DNA interaction by mwPIFE. ( A ) Scheme of DNA probes for detecting BamHI binding. Position of the BamHI restriction site is indicated. ( B ) Normalized PIFE values obtained with different probes incubated with BSA or BamHI. Error bars indicate standard deviations from measurements of three independent wells. P value was calculated by two-tailed t-test. ( C ) Effect of divalent cations on BamHI binding and cleavage. A 25 bp oligonucleotide with the restriction site 1 bp from Cy3 was used as a probe. Chart shows normalized PIFE values obtained upon addition of BamHI and after the subsequent wash. Error bars indicate standard deviations from measurements of three independent wells.

Article Snippet: After the scan, the buffer was replaced with 100 ul of a buffer containing appropriate protein BamHI (50 U), BSA (15 pmol, Biorad), Ku (0.5–20 pmol) or XPF/ERCC1 (25 pmol).

Techniques: Binding Assay, Incubation, Two Tailed Test

Journal: Nanoscale Research Letters

Article Title: Nanodiamonds as Carriers for Address Delivery of Biologically Active Substances

doi: 10.1007/s11671-010-9526-0

Figure Lengend Snippet: Radioactivity registered after chromatography of serum samples containing the RAM–nanodiamond–BSA I125 complex: 1 —sorbent with immobilized BSA, 2 —sorbent with immobilized mouse IgG (percentage of radioactivity of the initial samples used for chromatography)

Article Snippet: Mouse immunoglobulin G (IgG) (Medix Biochemical, Finland); bovine blood serum (Mikrogen, Russia); BSA (Amersham , England); Sepharose 6B (Pharmacia, Sweden); Rabbit AntiMouse Antibody (RAM) (Imtek Ltd., Russia); NaI125 (RITVERC, Russia); chloramine-T and sodium metabisulphite (Amersham , England) were used as acquired.

Techniques: Radioactivity, Chromatography

Journal: Cell Death and Differentiation

Article Title: A novel role for the apoptosis inhibitor ARC in suppressing TNFα-induced regulated necrosis

doi: 10.1038/cdd.2013.195

Figure Lengend Snippet: The ARC CARD is required for the ability of ARC to suppress TNF α -induced necrosis. ( a ) Immunoblot showing inhibition of TNF α -induced HMGB1 release by wild-type ARC but not by the CARD-defective double-point ARC mutant (L31F; G69R). L929 cells were stably transduced with empty vector (Φ), ARC-HA (ARC) or ARC-HA double-point CARD mutant (DM). Immunblot of bovine serum albumin (BSA) in media used as loading control. ( b ) Inhibition of TNF α -induced LDH release by ARC requires the CARD. Data shown as mean±S.E. from n =4. *** P -value

Article Snippet: Primary antibodies include: ARC (Cayman Chemical, Ann Arbor, MI, USA); β -actin (Sigma-Aldrich, St. Louis, MO, USA); HA (Roche Applied Science, Indianapolis, IN, USA); BSA, HMGB1, His (Abcam, Cambridge, MA, USA); histone H3, PARP, p65 (Cell Signaling Technology, Danvers, MA, USA); RhoGDI α (A-20) (Santa Cruz Biotechnology, Dallas, TX, USA); mouse RIP3 (Prosci, Poway, CA, USA); human RIP3 (Thermo Scientific); RIP1 (BD Biosciences, San Jose, CA, USA); and FADD (Enzo Life Sciences, Farmingdale, NY, USA).

Techniques: Inhibition, Mutagenesis, Stable Transfection, Transduction, Plasmid Preparation

Journal: Biomicrofluidics

Article Title: Acetylated bovine serum albumin differentially inhibits polymerase chain reaction in microdevices

doi: 10.1063/1.4983692

Figure Lengend Snippet: Effect of BSA type on the TaqMan-based PCR yield. Acetylated BSA from Ambion and Promega, non-acetylated BSA from Sigma were tested at 1 μ g/ μ l concentration in polypropylene tubes. Diamond illustrates a sample mean (center line) and 95% confidence interval (height).

Article Snippet: The PCR solution (20 μ l) contained 1× LuminoCt qPCR ReadyMix (Sigma Aldrich, Singapore), 1× ROX internal reference dye (Sigma Aldrich, Singapore), TaqMan probe for the nuc gene at 250 nM, forward and reverse primers at 900 nM each, 1 μ g/ μ l of either acetylated BSA (from two vendors: Ambion AM2614 and Promega P/N R3961) or non-acetylated BSA (Sigma-Aldrich P/N B8667), and 1 ng/ μ l of genomic DNA isolated from Staphylococcus aureus subsp. aureus ATCC 33591 (MRSA, strain 328; ATCC).

Techniques: Polymerase Chain Reaction, Concentration Assay