Journal: The American Journal of Pathology

Article Title: Elevated Peripheral Myelin Protein 22, Reduced Mitotic Potential, and Proteasome Impairment in Dermal Fibroblasts from Charcot-Marie-Tooth Disease Type 1A Patients

doi: 10.1016/j.ajpath.2017.10.021

Figure Lengend Snippet: List of Antibodies Used in This Study

Article Snippet: Rabbit , Ki67 , EnCor Biotechnology Inc. , RPCA-Ki67 , NA , 1:2000.

Techniques:

Journal: The American Journal of Pathology

Article Title: Elevated Peripheral Myelin Protein 22, Reduced Mitotic Potential, and Proteasome Impairment in Dermal Fibroblasts from Charcot-Marie-Tooth Disease Type 1A Patients

doi: 10.1016/j.ajpath.2017.10.021

Figure Lengend Snippet: Growth characteristics and cellular morphology of dermal fibroblasts from CMT1A patients. A and B: Quantification of the percentage of cells with incorporated BrdU ( A ), or Ki67 ( B ) reactivity in cultures from control individuals (C1–C4) and CMT1A patients (P1–P4). C: Representative micrographs of anti-vimentin immunofluorescence of controls (C3 and C4) and CMT1A patient (P1 to P4) fibroblasts, shown in red. All cells display elongated, kite-like morphology that is characteristic of dermal fibroblasts. Data are expressed as means ± SEM. * P < 0.05, *** P < 0.001 ( t -test). Scale bars = 20 μm.

Article Snippet: Rabbit , Ki67 , EnCor Biotechnology Inc. , RPCA-Ki67 , NA , 1:2000.

Techniques: Immunofluorescence

Journal: NPJ Parkinson's Disease

Article Title: TNFα increases tyrosine hydroxylase expression in human monocytes

doi: 10.1038/s41531-021-00201-x

Figure Lengend Snippet: a – d TH is detectable in recombinant form and in PC12 crude lysate using affinity-purified rabbit polyclonal TH antibody AB152 (Sigma), and antibodies selected for this ELISA, mouse monoclonal MCA-4H2 (EnCor), and rabbit polyclonal RPCA-TH (EnCor). e Using AB152, we probed the lower threshold for TH detection via serial dilution of purified recombinant TH from 6 to 0.094 μg/mL followed by Western blot and near-Infrared detection, considered to be a sensitive method for protein detection on Western blot. We demonstrate IR detection is reliable to a lower threshold of ~15 ng TH. Below this limit, TH detection becomes unreliable with IR detection. f – h In a series of stepwise experiments designed to increase ELISA sensitivity and decrease background, we achieved lower detection limits of 15 pg/mL TH ( h ). Capture antibody and detection antibody in all three methods were MCA-4H2 (1:1000 dilution from 1 mg/mL) and RPCA-TH (1:6000 dilution from 1 mg/mL). Schematic representation of each method shown on the left with a representative standard curve on the right. f Incubation with detection antibody followed by an HRP-conjugate secondary yielded a lower detection threshold of 125 pg/mL. g Addition of a tertiary layer using anti-rabbit biotin followed by Avidin-HRP improved lower detection threshold to 62.5 pg/mL but resulted in increased background. h Use of the biotinylated detection antibody (RPCA-TH-biotin, 1:6000 dilution from 1.65 mg/mL) followed by avidin-HRP yielded the lowest detection threshold of 15 pg/mL, with maximum sensitivity and minimal background. f – h Insets (red outline) shows a magnified lower standard curve to illustrate sensitivity.

Article Snippet: TH , Polyclonal/Rabbit , Biotin , EnCor , RPCA-TH , ELISA , 1:6000 , 1.65.

Techniques: Recombinant, Affinity Purification, Enzyme-linked Immunosorbent Assay, Serial Dilution, Purification, Western Blot, Incubation, Avidin-Biotin Assay

Journal: NPJ Parkinson's Disease

Article Title: TNFα increases tyrosine hydroxylase expression in human monocytes

doi: 10.1038/s41531-021-00201-x

Figure Lengend Snippet: a Schematic layout of experimental conditions to assess absorption controls in contrast to optimized Bio-ELISA conditions using PC12 cell lysate. b Representative standard curve shown to illustrate PC12 cells’ TH concentration using optimized Bio-ELISA (blue arrow), absorbed capture antibody (MCA-4H2 preincubated with 20 µg/mL recombinant TH, orange arrow), and absorbed detection antibody (biotinylated RPCA-TH preincubated with 20 µg/mL recombinant TH, green arrow). PC12 TH is undetectable after absorption of either capture or detection antibodies, confirming assay specificity.

Article Snippet: TH , Polyclonal/Rabbit , Biotin , EnCor , RPCA-TH , ELISA , 1:6000 , 1.65.

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

Journal: NPJ Parkinson's Disease

Article Title: TNFα increases tyrosine hydroxylase expression in human monocytes

doi: 10.1038/s41531-021-00201-x

Figure Lengend Snippet: Reagents and Materials.

Article Snippet: TH , Polyclonal/Rabbit , Biotin , EnCor , RPCA-TH , ELISA , 1:6000 , 1.65.

Techniques: Concentration Assay, Western Blot, Enzyme-linked Immunosorbent Assay, Plasmid Preparation, Isolation, Protease Inhibitor, Lysis, DC Protein Assay, Cell Culture, Cell Isolation, In Vitro

Journal: NPJ Parkinson's Disease

Article Title: TNFα increases tyrosine hydroxylase expression in human monocytes

doi: 10.1038/s41531-021-00201-x

Figure Lengend Snippet: Antibodies.

Article Snippet: TH , Polyclonal/Rabbit , Biotin , EnCor , RPCA-TH , ELISA , 1:6000 , 1.65.

Techniques: Concentration Assay, Enzyme-linked Immunosorbent Assay, Isolation, Avidin-Biotin Assay, Plasmid Preparation