Journal: Glycobiology

Article Title: Glycoproteomics enabled by tagging sialic acid- or galactose-terminated glycans

doi: 10.1093/glycob/cws144

Figure Lengend Snippet: GAL efficiently labels Gal and GalNAc residues on cell surface glycoproteins. ( A ) Asialo-K20 cells were incubated in PBS with 5 mg/mL BSA, pH 6.7, containing 250 μM aminooxy-biotin in the presence or absence of 10 mM aniline and 50 U/mL galactose

Article Snippet: For the oxime ligation step, cells were suspended to 1 × 107 cells/mL in PBS/5% BSA, pH 6.7 containing 250 μM aminooxy-biotin, aminooxy-AF488 (Invitrogen Corporation) or aminooxy-FLAG peptide and 10 mM aniline (Sigma–Aldrich) for 90 min at 4°C with end-over-end mixing.

Techniques: Incubation

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: 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: 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