Journal: Scientific Reports

Article Title: A method for reproducible measurements of serum BDNF: comparison of the performance of six commercial assays

doi: 10.1038/srep17989

Figure Lengend Snippet: BDNF ELISA Kits.

Article Snippet: Serum levels of total BDNF were measured by using six different ELISA kits, as listed in : human BDNF ELISA Kit (Cat #: SK00752-01, Aviscera-Bioscience, Santa Clara, CA, USA), BDNF Rapid TM ELISA Kit: Human, Mouse, Rat (2 Plates; Cat #: BEK-2211-2P, Biosensis Pty Ltd., SA, Australia), ChemKine TM BDNF Sandwich ELISA (Cat #: CYT306) and Milliplex ® Map Human Pituitary Magnetic Bead Panel 2 - Endocrine Multiplex Assay, based on Luminex ® /xMAP ® technology (Cat #: HPTP2MAG-66K, both from EMD Millipore Corporation, Billerica, MA, USA), BDNF Emax ® Immuno-Assay System (Cat #: G7610, Promega Corporation, Madison, WI, USA), Quantikine ® human BDNF Immunoassay (Cat #: DBD00, R&D Systems, Inc., Minneapolis, MN, USA).

Techniques: Enzyme-linked Immunosorbent Assay, Immuno Assay, Sandwich ELISA, Luminex, Recombinant, Magnetic Beads

Journal: Scientific Reports

Article Title: A method for reproducible measurements of serum BDNF: comparison of the performance of six commercial assays

doi: 10.1038/srep17989

Figure Lengend Snippet: Box plot of serum BDNF concentrations (ng/ml) from healthy volunteers (n = 38–40, see ) represented as mean of two independent measures. The upper line of the box marks the 75th percentile, the middle line is the median value and the lower line specifies the 25th percentile. Whiskers above and below the box indicate the 90th and 10th percentiles, respectively. Dots indicate the outlier values within each group.

Article Snippet: Serum levels of total BDNF were measured by using six different ELISA kits, as listed in : human BDNF ELISA Kit (Cat #: SK00752-01, Aviscera-Bioscience, Santa Clara, CA, USA), BDNF Rapid TM ELISA Kit: Human, Mouse, Rat (2 Plates; Cat #: BEK-2211-2P, Biosensis Pty Ltd., SA, Australia), ChemKine TM BDNF Sandwich ELISA (Cat #: CYT306) and Milliplex ® Map Human Pituitary Magnetic Bead Panel 2 - Endocrine Multiplex Assay, based on Luminex ® /xMAP ® technology (Cat #: HPTP2MAG-66K, both from EMD Millipore Corporation, Billerica, MA, USA), BDNF Emax ® Immuno-Assay System (Cat #: G7610, Promega Corporation, Madison, WI, USA), Quantikine ® human BDNF Immunoassay (Cat #: DBD00, R&D Systems, Inc., Minneapolis, MN, USA).

Techniques:

Journal: Scientific Reports

Article Title: A method for reproducible measurements of serum BDNF: comparison of the performance of six commercial assays

doi: 10.1038/srep17989

Figure Lengend Snippet: BDNF ELISA kits performance.

Article Snippet: Serum levels of total BDNF were measured by using six different ELISA kits, as listed in : human BDNF ELISA Kit (Cat #: SK00752-01, Aviscera-Bioscience, Santa Clara, CA, USA), BDNF Rapid TM ELISA Kit: Human, Mouse, Rat (2 Plates; Cat #: BEK-2211-2P, Biosensis Pty Ltd., SA, Australia), ChemKine TM BDNF Sandwich ELISA (Cat #: CYT306) and Milliplex ® Map Human Pituitary Magnetic Bead Panel 2 - Endocrine Multiplex Assay, based on Luminex ® /xMAP ® technology (Cat #: HPTP2MAG-66K, both from EMD Millipore Corporation, Billerica, MA, USA), BDNF Emax ® Immuno-Assay System (Cat #: G7610, Promega Corporation, Madison, WI, USA), Quantikine ® human BDNF Immunoassay (Cat #: DBD00, R&D Systems, Inc., Minneapolis, MN, USA).

Techniques: Enzyme-linked Immunosorbent Assay, Intra Assay, Inter Assay

Journal: Scientific Reports

Article Title: A method for reproducible measurements of serum BDNF: comparison of the performance of six commercial assays

doi: 10.1038/srep17989

Figure Lengend Snippet: Scatter plot showing the BDNF values distribution measured by the same operator on two different days using two plates of the same lot for each brand (Day 1 & Day 2). Each dot represents a BDNF value from one subject and the dashed lines link together two assessments of the same subject. The reproducibility was checked performing one-way ANOVA for repeated measures and the P values are specified.

Article Snippet: Serum levels of total BDNF were measured by using six different ELISA kits, as listed in : human BDNF ELISA Kit (Cat #: SK00752-01, Aviscera-Bioscience, Santa Clara, CA, USA), BDNF Rapid TM ELISA Kit: Human, Mouse, Rat (2 Plates; Cat #: BEK-2211-2P, Biosensis Pty Ltd., SA, Australia), ChemKine TM BDNF Sandwich ELISA (Cat #: CYT306) and Milliplex ® Map Human Pituitary Magnetic Bead Panel 2 - Endocrine Multiplex Assay, based on Luminex ® /xMAP ® technology (Cat #: HPTP2MAG-66K, both from EMD Millipore Corporation, Billerica, MA, USA), BDNF Emax ® Immuno-Assay System (Cat #: G7610, Promega Corporation, Madison, WI, USA), Quantikine ® human BDNF Immunoassay (Cat #: DBD00, R&D Systems, Inc., Minneapolis, MN, USA).

Techniques:

Journal: Scientific Reports

Article Title: A method for reproducible measurements of serum BDNF: comparison of the performance of six commercial assays

doi: 10.1038/srep17989

Figure Lengend Snippet: Scatter plots show the BDNF values distribution measured by the same operator on three different days for each brand. Distributions for Day 1 and Day 2 are the same as , here reported for comparison; distributions for Day 3 were obtained 1 year after Day 2, using a plate of a different lot (serum samples were stored at −80 °C). The reproducibility was checked performing one-way ANOVA for repeated measures and post-hoc Bonferroni correction applied when a significant comparison was found. P values and cumulative CVs expressed in percentage are given.

Article Snippet: Serum levels of total BDNF were measured by using six different ELISA kits, as listed in : human BDNF ELISA Kit (Cat #: SK00752-01, Aviscera-Bioscience, Santa Clara, CA, USA), BDNF Rapid TM ELISA Kit: Human, Mouse, Rat (2 Plates; Cat #: BEK-2211-2P, Biosensis Pty Ltd., SA, Australia), ChemKine TM BDNF Sandwich ELISA (Cat #: CYT306) and Milliplex ® Map Human Pituitary Magnetic Bead Panel 2 - Endocrine Multiplex Assay, based on Luminex ® /xMAP ® technology (Cat #: HPTP2MAG-66K, both from EMD Millipore Corporation, Billerica, MA, USA), BDNF Emax ® Immuno-Assay System (Cat #: G7610, Promega Corporation, Madison, WI, USA), Quantikine ® human BDNF Immunoassay (Cat #: DBD00, R&D Systems, Inc., Minneapolis, MN, USA).

Techniques: Comparison

Journal: Scientific Reports

Article Title: A method for reproducible measurements of serum BDNF: comparison of the performance of six commercial assays

doi: 10.1038/srep17989

Figure Lengend Snippet: ( A ) The antibodies from each ELISA kit were tested for specificity against pro-BDNF or mature BDNF. The BDNF standards blotted were commercial pro-BDNF (Alomone; 10 pg/lane), mature BDNF (1 and 2 from Alomone and Sigma, respectively; both 1000 pg/lane) and the standard BDNF protein included in each kit (Aviscera-Bioscience and Biosensis: 10 pg/lane; Millipore-ChemiKine TM , Millipore-Milliplex ® - and R&D System-Quantikine ® : 100 pg/lane; Promega-Emax ® : 1000 pg/lane). BSA (1000 pg/lane) was used as a negative control. The mouse monoclonal anti-BDNF antibody, (1:1000; Sigma) was tested as a control. ( B ) Central region of the same blot shown in A, from an overexposed film to better visualize the reactivity against pro-BDNF. ( C ) Reactivity of antibodies from Biosensis, Promega-Emax ® pAb and Sigma on a dot blot in which the same quantity of pro-BNDF and mature BDNF were spotted (100 pg each). Each antibody from the ELISA kits was used at the dilution suggested by the manufacturer’s instructions. mAb: Promega-Emax ® monoclonal capture antibody for plate coating. pAb: Promega-Emax ® polyclonal detection antibody.

Article Snippet: Serum levels of total BDNF were measured by using six different ELISA kits, as listed in : human BDNF ELISA Kit (Cat #: SK00752-01, Aviscera-Bioscience, Santa Clara, CA, USA), BDNF Rapid TM ELISA Kit: Human, Mouse, Rat (2 Plates; Cat #: BEK-2211-2P, Biosensis Pty Ltd., SA, Australia), ChemKine TM BDNF Sandwich ELISA (Cat #: CYT306) and Milliplex ® Map Human Pituitary Magnetic Bead Panel 2 - Endocrine Multiplex Assay, based on Luminex ® /xMAP ® technology (Cat #: HPTP2MAG-66K, both from EMD Millipore Corporation, Billerica, MA, USA), BDNF Emax ® Immuno-Assay System (Cat #: G7610, Promega Corporation, Madison, WI, USA), Quantikine ® human BDNF Immunoassay (Cat #: DBD00, R&D Systems, Inc., Minneapolis, MN, USA).

Techniques: Enzyme-linked Immunosorbent Assay, Negative Control, Control, Dot Blot

Journal: Molecular cell

Article Title: The Role of N-α-acetyltransferase 10 Protein in DNA Methylation and Genomic Imprinting

doi: 10.1016/j.molcel.2017.08.025

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: Other Abs used are against 5mC (BI-MECY-0500, Eurogentec), Dnmt1 for IP in (ab87654, Abcam), Naa10p for western in figure S1A (sc-33820, Santa Cruz), H3K9me3 (ab8898, Abcam), H3K9ac (07–352, Millipore), β-tubulin (MAB3408, Millipore), Tet2 (mAb-179–050, Diagenoda), Zfp57 (ab45341, Abcam), Trim28 (ab10483, Abcam), Lamin B (sc-6217, Santa Cruz), Uhrf1 (#12387, Cell Signaling) and α-tubulin (T5168, Sigma).

Techniques: Recombinant, SYBR Green Assay, Protease Inhibitor, Magnetic Beads, DNA Extraction, Multiplex Assay, Gel Extraction, Western Blot, Real-time Polymerase Chain Reaction, Methylation Sequencing, Mutagenesis, Plasmid Preparation, Software, Functional Assay

Journal: Cell Chemical Biology

Article Title: Site-Specific Photo-Crosslinking Proteomics Reveal Regulation of IFITM3 Trafficking and Turnover by VCP/p97 ATPase

doi: 10.1016/j.chembiol.2020.03.004

Figure Lengend Snippet:

Article Snippet: Peroxidase-AffiniPure goat anti-rabbit IgG (H+L) antibody , Jackson ImmunoResearch , Cat#111-035-003; RRID: AB_2313567.

Techniques: Virus, Recombinant, Protease Inhibitor, Multiplex sample analysis, Transfection, Western Blot, Antibody Labeling, BIA-KA, Mutagenesis, Control, Cloning, Plasmid Preparation, Software, Magnetic Beads

Journal: Cell Chemical Biology

Article Title: Site-Specific Photo-Crosslinking Proteomics Reveal Regulation of IFITM3 Trafficking and Turnover by VCP/p97 ATPase

doi: 10.1016/j.chembiol.2020.03.004

Figure Lengend Snippet: K24 Ubiquitination Regulates IFITM3 Turnover, Localization, and Co-trafficking with Incoming Virus Particles (A) Analysis of turnover of IFITM3 and K24A mutant. HeLa cells expressing HA-IFITM3 or K24A mutant were treated with CHX (25 μg/mL) for the indicated times and lysed for anti-HA western blot analysis. (B) Quantification of IFITM3 levels normalized to tubulin levels shown in (A). Data are represented as mean ± SD, n = 3. (C) Immunofluorescence analysis of IFITM3 and K24A mutant. HeLa cells were transfected with indicated HA-IFITM3 construct and mCherry-LAMP1, and processed for immunofluorescence with an Alexa Fluor 488-conjugated anti-HA antibody. Scale bars, 10 μm. (D) Quantification of Pearson coefficients for the IFITM3-LAMP1 colocalization shown in (C). Data are represented as mean ± SD, n = 50 cells. ∗∗∗∗p < 0.0001 calculated by Student's t test. (E) Relative percentage of DiD-IAV particles colocalized with IFITM3 and IFITM3-K24A at the time of dequenching. HeLa IFITM2/3-KO cells expressing BODIPY-labeled IFITM3 were infected with DiD-IAV particles and monitored for DiD dequenching and IFITM3 trafficking by time-lapse imaging. Data are represented as mean ± SD of three independent experiments. ∗∗p < 0.01 calculated by Student's t test. (F) Antiviral activity of IFITM3 and K24A mutant. HEK293T cells expressing HA-IFITM3 or HA-IFITM3-K24A were infected with IAV (PR8/H1N1) at an MOI of 2.5 for 6 h. Cells were fixed and stained with anti-HA and anti-influenza NP antibodies to measure IFITM3 expression and virus infection, respectively, with flow cytometry. “Low HA” and “High HA” indicate cell populations expressing low and high levels of HA-IFITM3, respectively. Data are represented as mean ± SD of three independent experiments. ∗∗p < 0.01, n.s. indicates p > 0.05 calculated by Student's t test.

Article Snippet: The secondary antibodies HRP-conjugated goat anti-mouse IgG (115-035-003) and HRP-conjugated goat anti-rabbit IgG (111-035-003) were purchased from Jackson ImmunoResearch.

Techniques: Ubiquitin Proteomics, Virus, Mutagenesis, Expressing, Western Blot, Immunofluorescence, Transfection, Construct, Labeling, Infection, Imaging, Activity Assay, Staining, Flow Cytometry

Journal: Cell Chemical Biology

Article Title: Site-Specific Photo-Crosslinking Proteomics Reveal Regulation of IFITM3 Trafficking and Turnover by VCP/p97 ATPase

doi: 10.1016/j.chembiol.2020.03.004

Figure Lengend Snippet: Chemical Inhibition of VCP ATPase Activity with ML240 Impairs IFITM3 Turnover, Lysosomal Sorting, and Trafficking (A) Analysis of IFITM3 and ubiquitination levels upon treatment with ML240. HEK293T cells were transfected with HA-IFITM3 for 12 h, treated with DMSO or ML240 (2.5 μM) for another 12 h, and then lysed for western blot analysis. The red asterisks indicate IFITM3 ubiquitination bands with high molecular weights. (B) Analysis of IFITM3 turnover upon treatment with ML240. HeLa cells expressing HA-IFITM3 were treated with CHX (25 μg/mL) for the indicated times in the presence of DMSO or ML240 (2.5 μM) and lysed for western blot analysis. (C) Quantification of IFITM3 levels normalized to tubulin levels shown in (B). Data are represented as mean ± SD, n = 3. (D) Immunofluorescence analysis of HA-IFITM3 in the presence of bafilomycin A1 or ML240. HeLa cells were transfected with HA-IFITM3 and mCherry-LAMP1, treated with DMSO, bafilomycin A1 (100 ng/mL), or ML240 (2.5 μM) for 12 h, and processed for immunofluorescence with an Alexa Fluor 488-conjugated anti-HA antibody. Scale bars, 10 μm. The right panels show magnified squared regions in the corresponding left panels. White arrows indicate enlarged IFITM3-containing compartments. (E) Relative percentage of DiD-IAV particles colocalized with IFITM3 at the time of dequenching upon ML240 treatment. HeLa IFITM2/3-KO cells expressing BODIPY-labeled IFITM3 were infected with DiD-IAV particles, acutely treated with ML240 (1 μM), and monitored for DiD dequenching and IFITM3 trafficking by time-lapse imaging. Data are represented as mean ± SD of three independent experiments. ∗∗∗∗p < 0.0001 calculated by Student's t test. (F) Antiviral activity of IFITM3 in the presence of ML240. HeLa IFITM2/3-KO cells were transfected with HA-IFITM3 and infected with IAV (PR8/H1N1) at an MOI of 2.5 for 2 h. Cells were then treated with ML240 (2.5 μM) for 4 h, fixed, and stained with anti-HA and anti-influenza NP antibodies to measure IFITM3 expression and virus infection, respectively, with flow cytometry. “Low HA” and “High HA” indicate cell populations expressing low and high levels of HA-IFITM3, respectively. Data are represented as mean ± SD of three independent experiments. ∗∗∗p < 0.001, n.s. indicates p > 0.05 calculated by Student's t test.

Article Snippet: The secondary antibodies HRP-conjugated goat anti-mouse IgG (115-035-003) and HRP-conjugated goat anti-rabbit IgG (111-035-003) were purchased from Jackson ImmunoResearch.

Techniques: Inhibition, Activity Assay, Ubiquitin Proteomics, Transfection, Western Blot, Expressing, Immunofluorescence, Labeling, Infection, Imaging, Staining, Virus, Flow Cytometry

Journal: Cell Chemical Biology

Article Title: Site-Specific Photo-Crosslinking Proteomics Reveal Regulation of IFITM3 Trafficking and Turnover by VCP/p97 ATPase

doi: 10.1016/j.chembiol.2020.03.004

Figure Lengend Snippet:

Article Snippet: The secondary antibodies HRP-conjugated goat anti-mouse IgG (115-035-003) and HRP-conjugated goat anti-rabbit IgG (111-035-003) were purchased from Jackson ImmunoResearch.

Techniques: Virus, Recombinant, Protease Inhibitor, Multiplex sample analysis, Transfection, Western Blot, Antibody Labeling, BIA-KA, Mutagenesis, Control, Cloning, Plasmid Preparation, Software, Magnetic Beads

Journal: Cell Chemical Biology

Article Title: Site-Specific Photo-Crosslinking Proteomics Reveal Regulation of IFITM3 Trafficking and Turnover by VCP/p97 ATPase

doi: 10.1016/j.chembiol.2020.03.004

Figure Lengend Snippet:

Article Snippet: Secondary antibodies goat anti-mouse IgG (Jackson ImmunoResearch, 115-035-174) and mouse anti-rabbit IgG (Jackson ImmunoResearch, 211-032-171) specific to light chain and conjugated to HRP were used in western blot of co-immunoprecipitates.

Techniques: Virus, Recombinant, Protease Inhibitor, Multiplex sample analysis, Transfection, Western Blot, Antibody Labeling, BIA-KA, Mutagenesis, Control, Cloning, Plasmid Preparation, Software, Magnetic Beads

Journal: Cell reports

Article Title: Neuronal activation of G αq EGL-30/GNAQ late in life rejuvenates cognition across species

doi: 10.1016/j.celrep.2023.113151

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: All secondary antibodies were used at a 1:500 dilution and included the following: Anti-Mouse IgG (H+L), highly cross-adsorbed, CF 647 antibody (Sigma-Aldrich, Cat. # SAB4600176), Anti-Rabbit IgG (H+L), highly cross-adsorbed, CF 555 antibody (Sigma-Aldrich, Cat. # SAB4600061), Alexa Fluor 647-AffiniPure Donkey Anti-Guinea Pig IgG (H+L) (Jackson ImmunoResearch, Cat. #706-605-148).

Techniques: Virus, Recombinant, Transfection, Magnetic Beads, Luciferase, RNAscope, Multiplex Assay, Reverse Transcription, Control, Plasmid Preparation, Software

Journal: Molecular cell

Article Title: The Role of N-α-acetyltransferase 10 Protein in DNA Methylation and Genomic Imprinting

doi: 10.1016/j.molcel.2017.08.025

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: Streptavidin HRP-conjugated , Jackson ImmunoResearch , 016-030-084; RRID: AB_2337238.

Techniques: Virus, Recombinant, Reverse Transcription, SYBR Green Assay, Protease Inhibitor, Magnetic Beads, DNA Extraction, Multiplex Assay, Gel Extraction, Western Blot, Real-time Polymerase Chain Reaction, Methylation Sequencing, Mutagenesis, Plasmid Preparation, Software, Functional Assay

Journal: iScience

Article Title: Methods to separate nuclear soluble fractions reflecting localizations in living cells

doi: 10.1016/j.isci.2021.103503

Figure Lengend Snippet: Key resources table

Article Snippet: donkey anti-goat IgG-Cy2 , Jackson ImmunoResearch Laboratories , Cat# 705-225-147; RRID: AB_2307341.

Techniques: Recombinant, Protease Inhibitor, Plasmid Preparation, Magnetic Beads, Bicinchoninic Acid Protein Assay, Multiplex sample analysis, Multiplex Assay, Extraction, Software, Microscopy