rabbit polyclonal anti-prph  (Millipore)

Journal: bioRxiv

Article Title: Astrocytes regulate neuronal network burst frequency through NMDA receptors species- and donor-specifically

doi: 10.1101/2023.12.09.570906

Figure Lengend Snippet: Characterization of hiPSC-derived neuron-astrocyte co-cultures. A. Differentiation of neurons and astrocytes. B, J-K. 5-week-old neurons expressed neuronal marker MAP2 and superficial cortical layer marker CUX1. C-D, L-M. The cultures did not stain for PRPH and PAX6, indicating the absence of peripheral neurons and differentiation-resistant PAX6-positive NPCs. E. The neurons expressed excitatory synaptic marker VGLUT1. F, N. There were no GAD67-positive cells in the cultures indicating the absence of GABAergic neurons. G-H, O-P. Approximately 50% of the cells in the co-cultures were S100β-positive astrocytes and 8-20-30% of the cells expressed astroglial marker GFAP. I. Neurons expressed co-localized Synapsin and PSD95 proteins for juxtaposed pre- and postsynaptic puncta, respectively. This indicated the presence of structural synapses. (n = 5 cell lines, data was collected from 3 independent experiments)

Article Snippet: Primary antibodies against MAP2 chicken (1:500, Abcam, Ab92434), CUX1 mouse (1:500, Abcam, ab54583), VGLUT1 rabbit (1:300, Sigma, vo389-200), GAD67 mouse (1:500, Abcam, ab26116), PRPH rabbit (1:2000, Novus, NB300-137), PAX6 rabbit (1:500, Thermo, 42-6600), Synapsin Oyster 650 mouse (1:500, Synaptic Systems, 106011C5), PSD95 rabbit (1:500, Cell Signaling Technologies, 3450), GRIN1 rabbit (1:500, Cell Signaling Technologies, 5704), s100β rabbit (1:100,Abcam, 52642) and GFAP rabbit (1:500, DAKO, Z033429-2) were used.

Techniques: Derivative Assay, Marker, Staining

Journal: Developmental Neurobiology

Article Title: Neuronal development in the cochlea of a nonhuman primate model, the common marmoset

doi: 10.1002/dneu.22850

Figure Lengend Snippet: Antibodies used in this study

Article Snippet: anti‐PRPH , rabbit , IgG , AB1530 , Merck Millipore, Burlington, MA, USA , 1:100.

Techniques:

Journal: Scientific Reports

Article Title: Selective Induction of Human Autonomic Neurons Enables Precise Control of Cardiomyocyte Beating

doi: 10.1038/s41598-020-66303-3

Figure Lengend Snippet: Human pluripotent stem cell (hPSC)-derived progenitors of the autonomic nervous system (ANS) differentiate into functional ANS neurons. (a ) Phase-contrast images of differentiated neurons on day 52. We induced hPSC-derived autonomic progenitors to mature neurons, using the autonomic neural crest (aNC) method shown in Fig. . Scale bar; left 100 μm, right 10 μm. ( b ) Distinct axon elongation of differentiated neurons on day 36. Neurons were cultured in the microfabricated device and labelled with a Synapsin-1-GFP lentiviral vector to detect axons. Neurons plated on the left-side chamber extended axons to the right-side chamber via microtunnels of 50-μm width (left panel). A highly magnified image of the boxed region in the middle panel is shown (right). ( c ) Immunofluorescence staining for TUJ1, PRPH, and TH on day 36. ( d) Comparison of the ratio of TH-positive to TUJ1-positive cells ( n = 5, error bar shows SDs; two-sided Student’s unpaired t -test * P < 0.001). The ratio of TH-positive areas to TUJ1-positive areas in induced neurons was calculated. ( e) Immunofluorescence staining of DBH and PRPH for sympathetic-like neurons on day 68 and PHOX2B and CHAT for parasympathetic-like neurons on day 52. Double immunofluorescence staining for TH and CHAT on day 59 were also shown. ( f ) Typical traces of calcium transients of induced neurons with indicated drugs. We used sensory neuron agonists menthol and capsaicin and ANS agonist nicotine. The right panel shows representative traces of calcium transients in 5 cells as shown in the phase-contrast image (left panel) for 350 s (see also Supplementary Movie ). The colour bar shows the fluorescent intensity. ( g ) Representative electrical signal in a selected single electrode after illumination. ChR2-expressing neurons were cultured on MEA substrates, and electrical activity was recorded. The left panel shows a schematic representation of the experimental protocol. Middle panels show phase-contrast and fluorescence microscopic images of neurons plated on MEA substrate. Blue lines indicate the timing of blue light stimulation in the right panel. Scale bar; 100 μm in ( b,c,e–g ).

Article Snippet: The following primary antibodies were used: mouse anti-NGFR (1:200; Advanced Targeting Systems, San Diego, CA, USA), mouse anti-class III beta-tubulin (TUJ1; 1:1000; Abcam, Cambridge, UK), mouse anti-microtubule-associated protein (MAP2; 1:1000; Abcam), mouse anti-PHOX2A (1:50; Abcam), mouse anti-PHOX2B (1:100; Proteintech, Rosemont, IL, USA), mouse anti-tyrosine hydroxylase (TH; 1:200; Merck Millipore, Billerica, MA, USA), mouse anti-dopamine β-hydroxylase (DBH; 1:50; Santa Cruz Biotechnology, Dallas, TX, USA), mouse anti-phenylethanolamine N -methyltransferase (PNMT; 1:100; Abcam), rabbit anti-SOX10 (1:500; Abcam), rabbit anti-Peripherin (PRPH; 1:1000; Merck Millipore), rabbit anti-TH (1:500; Merck Millipore), rabbit anti-PHOX2B (1:100; Abcam), and rabbit anti-choline acetyltransferase (CHAT; 1:1000; Abcam).

Techniques: Derivative Assay, Functional Assay, Cell Culture, Plasmid Preparation, Immunofluorescence, Staining, Double Immunofluorescence Staining, Expressing, Activity Assay, Fluorescence

Journal: Scientific Reports

Article Title: Selective Induction of Human Autonomic Neurons Enables Precise Control of Cardiomyocyte Beating

doi: 10.1038/s41598-020-66303-3

Figure Lengend Snippet: Cell density and neurotrophic factor facilitate selective differentiation of sympathetic-like and parasympathetic-like neurons. ( a ) Single-cell RNA-seq analysis using differentiated cells on day 58. The t-distributed stochastic neighbor embedding (t-SNE) visualization showing 5 distinct cell populations (clusters A–E) based on K-means clustering. ( b ) Violin plots indicating the expression levels of STMN2 and PRPH across five clusters. ( c ) Expression patterns of peripheral neuron markers, sympathetic neuron markers, and parasympathetic neuron markers in each neuron of cluster D and E. ( d ) Immunofluorescence analysis of 9 culture conditions. Scale bar; 100 μm. ( e ) Expression fold-changes in TH-positive and CHAT-positive neurons under 9 culture conditions. Each result is normalized to the positive region in the cc1 condition ( n = 3; error bar shows SDs; one-way ANOVA Tukey post-test * P < 0.05 compared to cc1 for TH-positive neurons. one-way ANOVA Tukey post-test † P < 0.05 compared to r1 condition for CHAT-positive neurons). ( f ) mRNA expression levels of PHOX2B, TH, and CHAT under each culture condition ( n = 3; error bar shows SDs; two-sided Student’s unpaired t -test for PHOX2B and CHAT. Welch’s t -test for TH. * P < 0.01, ** P < 0.05).

Article Snippet: The following primary antibodies were used: mouse anti-NGFR (1:200; Advanced Targeting Systems, San Diego, CA, USA), mouse anti-class III beta-tubulin (TUJ1; 1:1000; Abcam, Cambridge, UK), mouse anti-microtubule-associated protein (MAP2; 1:1000; Abcam), mouse anti-PHOX2A (1:50; Abcam), mouse anti-PHOX2B (1:100; Proteintech, Rosemont, IL, USA), mouse anti-tyrosine hydroxylase (TH; 1:200; Merck Millipore, Billerica, MA, USA), mouse anti-dopamine β-hydroxylase (DBH; 1:50; Santa Cruz Biotechnology, Dallas, TX, USA), mouse anti-phenylethanolamine N -methyltransferase (PNMT; 1:100; Abcam), rabbit anti-SOX10 (1:500; Abcam), rabbit anti-Peripherin (PRPH; 1:1000; Merck Millipore), rabbit anti-TH (1:500; Merck Millipore), rabbit anti-PHOX2B (1:100; Abcam), and rabbit anti-choline acetyltransferase (CHAT; 1:1000; Abcam).

Techniques: RNA Sequencing Assay, Expressing, Immunofluorescence