Journal: eLife

Article Title: Chondroitin sulfate proteoglycan 4,6 sulfation regulates sympathetic nerve regeneration after myocardial infarction

doi: 10.7554/eLife.78387

Figure Lengend Snippet:

Article Snippet: Peptide/recombinant protein , NGF , Alomone Labs , N-100 , (10 ng/mL).

Techniques: Knockdown, Staining, Recombinant, Sequencing, In Vitro, In Vivo, Formulation, Software, Imaging

Journal: Frontiers in Pharmacology

Article Title: Jia-Wei-Kai-Xin-San, an Herbal Medicine Formula, Ameliorates Cognitive Deficits via Modulating Metabolism of Beta Amyloid Protein and Neurotrophic Factors in Hippocampus of Aβ 1-42 Induced Cognitive Deficit Mice

doi: 10.3389/fphar.2019.00258

Figure Lengend Snippet: Jia-Wei-Kai-Xin-San treatment induced transformation of neurotrophic factors precursors into mature forms in hippocampus of Aβ injection mice. (A) The treatment of JWKXS and huperzine A in the mice were same as that in . The hippocampus tissues were collected after 1-week treatment and the expressions of proNGF and NGF were analyzed by western blot analysis and ELISA kits. Afterward, the ratio of mNGF to proNGF was calculated. (B) Expressions of proBDNF and BDNF were determined and the ratio of BDNF to proBDNF was calculated as same as that of NGF. Comparisons between groups were carried out by a one-way ANOVA followed by a post hoc Bonferroni test. Values were expressed in the percentage of control group as Mean ± SEM ( n = 8). # p < 0.05 (compared with sham group); ∗ p < 0.05, ∗∗ p < 0.01 (compared with model group).

Article Snippet: Aβ amounts were determined by amyloid beta protein mouse ELISA Kit (Jinyibai Company, China). proNGF and NGF levels were determined by mouse proNGF ELISA kit and mouse NGF ELISA kit (CUSABIO, Houston, TX, United States). proBDNF and BDNF levels were determined by mouse proBDNF ELISA kit and mouse BDNF ELISA kit (Aviscera Bioscience, Santa Clara, CA, United States).

Techniques: Transformation Assay, Injection, Western Blot, Enzyme-linked Immunosorbent Assay, Control

Journal: Investigative and Clinical Urology

Article Title: Treatment with low-energy shock wave alleviates pain in an animal model of uroplakin 3A-induced autoimmune interstitial cystitis/painful bladder syndrome

doi: 10.4111/icu.2019.60.5.359

Figure Lengend Snippet: Systemic and local inflammation in bladder tissue was induced by uroplakin 3A (UPK3A) immunization, and low-energy shock wave (LESW) treatment decreased local inflammatory reactions. (A) Serum levels of tumor necrosis factor-α (TNF-α), nerve growth factor (NGF), interleukin (IL)-2, and IL-6, respectively, influenced by UPK3A immunization and LESW treatment. (B) Reverse transcription polymerase chain reaction examination of expression levels of NGF, interferon-γ (IFN-γ), TNF-α, and IL-2, -4, -6, and -8. (C) Western blot examination of protein levels of TNF-α, NGF, and downstream inflammatory marker nuclear factor-κB (NF-κB). (D) Calculation of Western blot results based on bands' grey scale. * p<0.05 compared to Sham. # p<0.05 compared to UPK3A.

Article Snippet: Concentrations of TNF-α (575209; Biolegend, San Diego, CA, USA), NGF (E-EL-M0815c; Elabscience, Houston, TX, USA), interleukin-2 (IL-2, 575409; Biolegend), and IL-6 (575709; Biolegend) were measured using ELISA kits according to the manufacturers' instructions.

Techniques: Reverse Transcription, Polymerase Chain Reaction, Expressing, Western Blot, Marker

Journal: Frontiers in Molecular Neuroscience

Article Title: NGF and proNGF Reciprocal Interference in Immunoassays: Open Questions, Criticalities, and Ways Forward

doi: 10.3389/fnmol.2016.00063

Figure Lengend Snippet: Recovery of NGF and proNGF spiked in the sample buffer (supplied by each supplier of the commercial kits) .

Article Snippet: The CUSABIO Mouse Pro-Nerve Growth Factor (proNGF) ELISA Kit was purchased and tested.

Techniques: Enzyme-linked Immunosorbent Assay

Journal: Frontiers in Molecular Neuroscience

Article Title: NGF and proNGF Reciprocal Interference in Immunoassays: Open Questions, Criticalities, and Ways Forward

doi: 10.3389/fnmol.2016.00063

Figure Lengend Snippet: NGF and proNGF spiked into brain cortex extract of proNGF#72 Transgenic Mouse assayed by Emax Promega with mAb Promega and mAb αD11 .

Article Snippet: The CUSABIO Mouse Pro-Nerve Growth Factor (proNGF) ELISA Kit was purchased and tested.

Techniques: Transgenic Assay, Concentration Assay, Standard Deviation

Journal: Frontiers in Molecular Neuroscience

Article Title: NGF and proNGF Reciprocal Interference in Immunoassays: Open Questions, Criticalities, and Ways Forward

doi: 10.3389/fnmol.2016.00063

Figure Lengend Snippet: IP and WB of transgenic and wild-type mice. (A): IP and WB of cortex extracts from male (M) and female (F) TgProNGF#72 and wild-type (WT) mice. IP on extracts from cortex (CTX) with anti-NGF αD11 antibody, followed by WB with anti-NGF or anti-proNGF antibody, as described in Tiveron et al. . A representative WB probed with anti-proNGF (PAb Alomone), (top) or anti-NGF M20 (Santa Cruz) (bottom) is shown. TgproNGF#72 and wild type mice, male and female, were analyzed. (B) Quantitative analysis of proNGF and mature NGF in the CTX of TgproNGF#72 mice, male and female, by IP and WB and densitometric analysis. After anti-NGF IP, the proNGF bands, (in WB probed with anti-proNGF), and the NGF bands, (in the WB probed with anti-NGF antibody), both identified also by Mass Spectrometry, were quantified. The resulting intensities were normalized against the area of the bands, and then compared with an internal standard of recombinant proNGF and NGF. Loaded samples were in the linear range of detection. Comparison between proNGF and NGF amounts in TgproNGF#72, male and female, is reported in the histogram. The experiment was carried out in triplicate.

Article Snippet: The CUSABIO Mouse Pro-Nerve Growth Factor (proNGF) ELISA Kit was purchased and tested.

Techniques: Transgenic Assay, Mass Spectrometry, Recombinant, Comparison

Journal: Frontiers in Molecular Neuroscience

Article Title: NGF and proNGF Reciprocal Interference in Immunoassays: Open Questions, Criticalities, and Ways Forward

doi: 10.3389/fnmol.2016.00063

Figure Lengend Snippet: NGF and proNGF spiked in the sample buffer (supplied by each supplier of the commercial kits) . The histograms summarize the results of the NGF and proNGF spiked separately and together into the assay buffer of the three different commercial kits for the detection of NGF analyzed. The Promega kit was tested in the two different formats: rat mAb Promega and αD11. Two concentrations of recombinant NGF (10 and 100 pg/ml) and proNGF (50 and 200 pg/ml) were spiked into the assay buffer either alone or together with NGF and measured by using the kits previously described. (A) (Emax Promega with mAb Promega), (B) (Emax Promega with mAb αD11), (C) (Chemikine Chemicon), (D) (NGF Rapid ELISA Biosensis) report the values of concentration, interpolated by the calibration curves. The calculated values of the samples spiked with both NGF and proNGF were compared to the spiked value of only NGF. The t -student test was carried out and the p -values were calculated. Two asterisks on the histograms mean a p < 0.001.

Article Snippet: The CUSABIO Mouse Pro-Nerve Growth Factor (proNGF) ELISA Kit was purchased and tested.

Techniques: Recombinant, Enzyme-linked Immunosorbent Assay, Concentration Assay

Journal: Frontiers in Molecular Neuroscience

Article Title: NGF and proNGF Reciprocal Interference in Immunoassays: Open Questions, Criticalities, and Ways Forward

doi: 10.3389/fnmol.2016.00063

Figure Lengend Snippet: ELISA for the differential detection of NGF and ProNGF . (A) Strategy: capture NGF and measure proNGF (Exploiting the mAb αD11 fast kinetics). ELISA sandwiches format based on a pre-capturing of NGF by a treatment of the sample with mAb αD11 (Cattaneo et al., ) on a solid support. Subsequent detection of proNGF by traditional sandwich ELISA. Antibodies tested in different combinations: Anti-NGF pAb H20 (Santa Cruz no. sc-548), Anti-proNGF scFV FPro10 (Paoletti et al., ), Anti-NGF pAb M20 (Santa Cruz no. sc-549), Anti-NGF pAb (Sigma no. N 6655), Anti-proNGF pAb (Sigma no. P 5498), mAb αD11 (Cattaneo et al., ), Anti-NGF mAb 256 (R&D no. MAB256). (B) Strategy: capture proNGF and measure proNGF (Exploiting anti—proNGF antibodies in ELISA sandwich). Antibodies tested in different combinations: Anti-proNGF scFV FPro10 (Paoletti et al., ), mAb αD11 (Cattaneo et al., ), Anti-NGF pAb M20 (Santa Cruz no. sc-549), Anti-proNGF Novus (no. S-080-100), Anti-NGF mAb 256 (R&D no. MAB256), Anti-proNGF mAb (clone EP1318Y) (Millipore no. 04-1142), Anti-proNGF pAb Chemicon (Millipore no. AB9040), Anti-proNGF pAb Alomone (no. ANT-005), Anti-NGF Abnova (no. PAB0755).

Article Snippet: The CUSABIO Mouse Pro-Nerve Growth Factor (proNGF) ELISA Kit was purchased and tested.

Techniques: Enzyme-linked Immunosorbent Assay, Sandwich ELISA

Journal: Frontiers in Molecular Neuroscience

Article Title: NGF and proNGF Reciprocal Interference in Immunoassays: Open Questions, Criticalities, and Ways Forward

doi: 10.3389/fnmol.2016.00063

Figure Lengend Snippet: SPR analysis—calibration curves obtained with proNGF and NGF over the panel of different antibodies. (A) Different proNGF concentrations tested on anti-proNGF mAb FPro10. Concentrations in nM (from top): 500, 333, 222, 148, 98, 65, 33, 16, 8, 4, 2, 1, 0.5; (B) Different proNGF concentrations tested on anti-proNGF mAb Millipore. Concentrations in nM (from top): 500, 333, 222, 148, 98, 65, 33, 16, 8, 4, 2, 1, 0.5; (C) Different proNGF concentrations tested on anti-NGF mAb αD11. Concentrations in nM (from top): 500, 333, 222, 148, 98, 65, 33, 16, 8, 4, 2, 1, 0.5; (D) Different proNGF concentrations tested on anti-NGF mAb R&D. Concentrations in nM (from top): 100, 50, 25, 12.50, 6.25, 3.2, 1.6, 0.8, 0.4, 0.2, 0.1; (E) Different NGF concentrations tested on anti-NGF mAb R&D. Concentrations in nM (from top): 100, 50, 25, 12.50, 6.25, 3.2, 1.6, 0.8, 0.4, 0.2, 0.1.

Article Snippet: The CUSABIO Mouse Pro-Nerve Growth Factor (proNGF) ELISA Kit was purchased and tested.

Techniques:

Journal: Frontiers in Molecular Neuroscience

Article Title: NGF and proNGF Reciprocal Interference in Immunoassays: Open Questions, Criticalities, and Ways Forward

doi: 10.3389/fnmol.2016.00063

Figure Lengend Snippet: Binding analysis of NGF and proNGF mixtures to different antibodies . The following antibodies were immobilized on the chip for SPR experiments. (A) anti-proNGF mAb FPro10; (B) anti-proNGF mAb Millipore; (C) anti-NGF mAb αD11. In all the panels, the curves represent the following analytes from top to bottom: 200 nM proNGF + 20 nM NGF (solid blue line, experimental value); 200 nM proNGF (solid yellow line, experimental value); 200 nM proNGF + 20 nM NGF (segmented blue line, theoretical value); 40 nM proNGF + 20 nM NGF (solid purple line, experimental value); 40 nM proNGF (solid orange line, experimental value); 40 nM proNGF + 20 nM NGF (segmented purple line, theoretical value); 20 nM proNGF + 20 nM NGF (solid green line, experimental value); 20 nM proNGF (solid red line, experimental value); 20 nM proNGF + 20 nM NGF (segmented green line, theoretical value); 20 nM NGF (solid dark blue line, experimental value). The segmented lines represents the theoretical curves for the point-to-point algebraic sum of the experimental curves of the single components, at the concentrations indicated.

Article Snippet: The CUSABIO Mouse Pro-Nerve Growth Factor (proNGF) ELISA Kit was purchased and tested.

Techniques: Binding Assay

Journal: Frontiers in Molecular Neuroscience

Article Title: NGF and proNGF Reciprocal Interference in Immunoassays: Open Questions, Criticalities, and Ways Forward

doi: 10.3389/fnmol.2016.00063

Figure Lengend Snippet: Calculations of the theoretical value of the measured RU for the single NGF or proNGF components and comparison to the experimental one .

Article Snippet: The CUSABIO Mouse Pro-Nerve Growth Factor (proNGF) ELISA Kit was purchased and tested.

Techniques: Comparison

Journal: Frontiers in Molecular Neuroscience

Article Title: NGF and proNGF Reciprocal Interference in Immunoassays: Open Questions, Criticalities, and Ways Forward

doi: 10.3389/fnmol.2016.00063

Figure Lengend Snippet: AlphaLISA experiments: curves of NGF, proNGF, mix of NGF + proNGF, comparison at different incubation times . Curves of NGF, proNGF, mix of NGF + proNGF, linearly interpolated. NGF and proNGF were assayed separately and together (dynamic range: 2000–4 pg/ml, 1:2 dilutions, in duplicates), in different stoichiometric ratio. The dilution were done in AlphaLISA NaCl buffer (Perkin Elmer). The protocol is described in the methods sections and called for two incubation times. In (A) the first incubation time lasted 60 min and the second one 30 min, as suggested by the manufacturer's protocol. In (B) , a comparison between different incubation times is shown. The first incubation time was 15, 30, or 60 min, while the second one was 20 min for all the curves. The signal was read on the Perkin Elmer instrument EnVision®.

Article Snippet: The CUSABIO Mouse Pro-Nerve Growth Factor (proNGF) ELISA Kit was purchased and tested.

Techniques: Comparison, Incubation

Journal: Frontiers in Molecular Neuroscience

Article Title: NGF and proNGF Reciprocal Interference in Immunoassays: Open Questions, Criticalities, and Ways Forward

doi: 10.3389/fnmol.2016.00063

Figure Lengend Snippet: ProNGF CUSABIO kit: serial dilution of the samples derived from brain extract of TgProNGF#72 and WT mice .

Article Snippet: The CUSABIO Mouse Pro-Nerve Growth Factor (proNGF) ELISA Kit was purchased and tested.

Techniques: Serial Dilution, Derivative Assay, Concentration Assay, Standard Deviation

Journal: Frontiers in Molecular Neuroscience

Article Title: NGF and proNGF Reciprocal Interference in Immunoassays: Open Questions, Criticalities, and Ways Forward

doi: 10.3389/fnmol.2016.00063

Figure Lengend Snippet: proNGF CUSABIO kit: Spiking of recombinant mouse proNGF into TgProNGF#72and WT mice brain samples .

Article Snippet: The CUSABIO Mouse Pro-Nerve Growth Factor (proNGF) ELISA Kit was purchased and tested.

Techniques: Recombinant, Standard Deviation

Journal: Frontiers in Molecular Neuroscience

Article Title: NGF and proNGF Reciprocal Interference in Immunoassays: Open Questions, Criticalities, and Ways Forward

doi: 10.3389/fnmol.2016.00063

Figure Lengend Snippet: Analysis of transgenic and wild-type mice samples by SPR . Injection of samples from transgenic and from WT mice over the FPro10 anti-proNGF antibody (A) and over the Millipore anti-proNGF antibody (B) (blank subtracted curves). Representation of the SPR curves after subtraction of the WT mice signals. Curve of TgproNGF#72 mice subtracted the curve of WT for the FPro10 anti-proNGF antibody ( A —insert), and for the Millipore anti-proNGF antibody ( B —insert). In all panels: Red curve: TgproNGF#72, blue curve: WT.

Article Snippet: The CUSABIO Mouse Pro-Nerve Growth Factor (proNGF) ELISA Kit was purchased and tested.

Techniques: Transgenic Assay, Injection

Journal: Molecular imaging and biology

Article Title: PET detection of cerebral necrosis using an infarct-avid agent 2-deoxy-2-[ 18 F]fluoro-D-glucaric acid (FGA) in a mouse model of the brain stroke

doi: 10.1007/s11307-020-01513-9

Figure Lengend Snippet: Concentration of neurotrophin levels in the brain. (A) BDNF levels (n=5). (B) NGF-β levels (n = 5 from stroke group and n = 6 from sham group).

Article Snippet: Brain-derived nerve factor (BDNF) and nerve growth factor (NGF)-β Mouse BDNF and NGF-β ELISA kits were purchased from Boster Biochem (Pleasanton, CA).

Techniques: Concentration Assay