rabbit anti-mouse trka  (Thermo Fisher)

Journal: Experimental eye research

Article Title: Analysis of 14-3-3 isoforms expressed in photoreceptors

doi: 10.1016/j.exer.2018.02.022

Figure Lengend Snippet: Antibodies & Lectins used for immunohistochemistry (IHC) or Western blot (WB)

Article Snippet: RRID Rabbit anti-pan 14-3-3 Abcam Ab9063 1:250 (IHC), 1:300 (WB) AB_306972 Mouse anti-pan 14-3-3 (H8) Santa Cruz sc-1657 1:100 (IHC), 1:500 (WB) AB_626618 Rabbit anti-14-3-3 epsilon Cell Signaling 9635 1:100 (IHC), 1:1000 (WB) n/a Mouse anti-14-3-3 zeta (1B3) Santa Cruz sc-293415 1:100 (IHC), 1:500 (WB) n/a WGA: Alexa 594 ThermoFisher W11262 1:500 (IHC) n/a Goat anti-rabbit IgG: Alexa 647 ThermoFisher A21244 1:500 (IHC) AB_141663 Goat anti-mouse IgG: Alexa 488 ThermoFisher A11001 1:500 (IHC) AB_2534069 IRDye ® 680RD Goat anti-Rabbit IgG LI-COR 925-68071 1:20000 (WB) n/a IRDye ® 800CW Goat anti-Mouse IgG LI-COR 925-32210 1:20000 (WB) n/a Open in a separate window Antibodies & Lectins used for immunohistochemistry (IHC) or Western blot (WB)

Techniques: Immunohistochemistry, Western Blot

Journal: Experimental eye research

Article Title: Analysis of 14-3-3 isoforms expressed in photoreceptors

doi: 10.1016/j.exer.2018.02.022

Figure Lengend Snippet: 14-3-3 proteins are expressed throughout the retina. A–B) mouse retina labeled with WGA (left), pan 14-3-3 (center, green), with merged images including Hoechst labeling for nuclei on the right. A polyclonal pan 14-3-3 antibody (Abcam) was used in A, and a monoclonal pan-14-3-3 (H8) in B. Asterisks in (B) indicate labeling of blood vessels by the secondary antibody. Scale bar, 20 μm and abbreviations are: OS, outer segment; IS, inner segment; ONL, outer nuclear layer; OPL, outer plexiform layer; INL, inner nuclear layer; IPL, inner plexiform layer; GC, ganglion cell layer. C) Western blots of recombinant 14-3-3 proteins probed with the antibodies used in A–B; blots stained with REVERT to label total protein. Quantitation of the band intensities, data represent mean + SD, reveals no statistically significant differences between the two antibodies.

Article Snippet: RRID Rabbit anti-pan 14-3-3 Abcam Ab9063 1:250 (IHC), 1:300 (WB) AB_306972 Mouse anti-pan 14-3-3 (H8) Santa Cruz sc-1657 1:100 (IHC), 1:500 (WB) AB_626618 Rabbit anti-14-3-3 epsilon Cell Signaling 9635 1:100 (IHC), 1:1000 (WB) n/a Mouse anti-14-3-3 zeta (1B3) Santa Cruz sc-293415 1:100 (IHC), 1:500 (WB) n/a WGA: Alexa 594 ThermoFisher W11262 1:500 (IHC) n/a Goat anti-rabbit IgG: Alexa 647 ThermoFisher A21244 1:500 (IHC) AB_141663 Goat anti-mouse IgG: Alexa 488 ThermoFisher A11001 1:500 (IHC) AB_2534069 IRDye ® 680RD Goat anti-Rabbit IgG LI-COR 925-68071 1:20000 (WB) n/a IRDye ® 800CW Goat anti-Mouse IgG LI-COR 925-32210 1:20000 (WB) n/a Open in a separate window Antibodies & Lectins used for immunohistochemistry (IHC) or Western blot (WB)

Techniques: Labeling, Western Blot, Recombinant, Staining, Protein Quantitation

Journal: Journal of Neuropathology & Experimental Neurology

Article Title: Hippocampal ProNGF Signaling Pathways and β-Amyloid Levels in Mild Cognitive Impairment and Alzheimer Disease

doi: 10.1097/nen.0b013e318272caab

Figure Lengend Snippet: FIGURE 1. Box plots and representative immunoblots of hippocampal levels of ProNGF, TrkA, p75NTR, and NRH2 in cases clinically diagnosed as NCI, MCI, and AD. Immunoreactive signals were normalized to A-tubulin levels on the same blots by densitometry. (A) Levels of proNGF were significantly elevated in AD versus NCI and MCI. * p = 0.004. (B) Levels of TrkA were significantly reduced in MCI and upregulated in AD. * p = 0.014. Levels of p75NTR (C) and NRH2 (D) remained stable across clinical diagnoses.

Article Snippet: The antibodies included proNGF polyclonal antiserum (1:50, H-20; Santa Cruz Biotechnology, Santa Cruz, CA); purified anti-TrkA rabbit polyclonal affinityYpurified antibodies (1:100; Fitzgerald, Acton, MA); anti-NRH2 (1:1000), antisortilin (1:1000), and anti-p75NTR (1:500) obtained from Abcam (Cambridge, MA); and anti-Akt (1:1000), phosphoAkt (Ser473) (1:1000), p44/p42 MAPK (Erk1/2) (1:1000), phospho-p44/p42 MAPK (Erk1/2) (Thr202/Tyr 204) (E10) mouse monoclonal antibody (mAb) (1:2000), SAPK/JNK (56G8) rabbit mAb (1:1000), and phospho-SAPK/JNK (Thr183/Tyr185) (81E11) rabbit mAb (1:2000) obtained from Cell Signaling Technology (Danvers, MA).

Techniques: Western Blot

Journal: Journal of Neuropathology & Experimental Neurology

Article Title: Hippocampal ProNGF Signaling Pathways and β-Amyloid Levels in Mild Cognitive Impairment and Alzheimer Disease

doi: 10.1097/nen.0b013e318272caab

Figure Lengend Snippet: FIGURE 5. Schematic summary diagram showing the relative changes for proNGF, TrkA, p75NTR, NRH2, Erk, Akt, phospho- Akt, JNK, and phospho-JNK in the hippocampus during the progression of AD. mAD, mild AD.

Article Snippet: The antibodies included proNGF polyclonal antiserum (1:50, H-20; Santa Cruz Biotechnology, Santa Cruz, CA); purified anti-TrkA rabbit polyclonal affinityYpurified antibodies (1:100; Fitzgerald, Acton, MA); anti-NRH2 (1:1000), antisortilin (1:1000), and anti-p75NTR (1:500) obtained from Abcam (Cambridge, MA); and anti-Akt (1:1000), phosphoAkt (Ser473) (1:1000), p44/p42 MAPK (Erk1/2) (1:1000), phospho-p44/p42 MAPK (Erk1/2) (Thr202/Tyr 204) (E10) mouse monoclonal antibody (mAb) (1:2000), SAPK/JNK (56G8) rabbit mAb (1:1000), and phospho-SAPK/JNK (Thr183/Tyr185) (81E11) rabbit mAb (1:2000) obtained from Cell Signaling Technology (Danvers, MA).

Techniques:

Journal: Nature neuroscience

Article Title: An evolving NGF–Hoxd1 signaling pathway mediates development of divergent neural circuits in vertebrates

doi: 10.1038/nn.2710

Figure Lengend Snippet: (a) Diagram of the numbers of unique genes identified by the three microarray screens (at a loose cutoff of probability > 0.5 and fold change > 1.5). (b) The degree of mRNA level changes of the 14 genes identified by all three independent microarray screens, in log scale. (c) Application of NGF to cultured mouse DRG explants robustly induces transcription of Hoxd1 within 24 hrs. (d) RT-PCR reactions comparing levels of Hoxd1 mRNA in DRGs dissected from e14.5 Bax −/− and Ngf −/− ; Bax −/− embryos. (e) In parallel cultures of mouse and chick DRG explants, NGF induces robust transcription of canonical NGF-dependent genes ( CGRP and Ret ) in both species, but induces Hoxd1 in mouse but not in chick DRGs as assessed by quantitative PCR. * p < 0.05 by Student’s t -test (n = 3 independent batches of cultures for each condition). (f) In situ hybridization for Hoxd1 mRNA in sections of developing DRGs of e14.5 Bax −/− and Ngf −/− ; Bax −/− embryos. Scale bar = 50 μm. (g) Comparison of Hoxd1 expression in DRGs and spinal cord of e15.5 mouse and stage 32 (e7.5) chick embryos. TrkA is a general marker for embryonic nociceptors. In chick DRGs, TrkA + nociceptors and TrkA non-nociceptors segregate into distinct topographic domains. sc, spinal cord. drg, dorsal root ganglion. Scale bar = 50 μm and 100 μm in low- and high-magnification panels, respectively. (h) Double fluorescent labeling of Hoxd1 mRNA and TrkA protein in e15.5 mouse and stage 32 chick DRGs. Scale bar = 20 μm.

Article Snippet: Primary antibodies used include: rabbit anti-mouse TrkA (Millipore, 1:1000), rabbit anti-chicken TrkA and TrkC (gifts from Dr. Frances Lefcort, Montana State University, 1:1000), rabbit (Millipore, 1:1000) or mouse anti-peripherin (Millipore, 1:500), rabbit anti-CGRP (Millipore, 1:1000), rabbit anti-PGP9.5 (Millipore, 1:1000), rabbit anti-Parvalbumin (Swant, 1:2000), rabbit anti-GFP (1:2000, Invitrogen), chicken anti-LacZ (Promega, 1:500), rabbit anti-Ret (Immuno-Biological Laboratories, 1:1000), mouse anti-chick Neurofilament 160kD (Zymed, 1:500) and rabbit anti-Neurofilament 200 (Millipore, 1:1000).

Techniques: Microarray, Cell Culture, Reverse Transcription Polymerase Chain Reaction, Real-time Polymerase Chain Reaction, In Situ Hybridization, Comparison, Expressing, Marker, Labeling

Journal: Nature neuroscience

Article Title: An evolving NGF–Hoxd1 signaling pathway mediates development of divergent neural circuits in vertebrates

doi: 10.1038/nn.2710

Figure Lengend Snippet: (a) Diagram showing the distinct patterns of nociceptor endings in the skin of mammals and birds. (b) Anti-peripherin staining of mouse hairy skin from proximal limb. Upper panels: transverse sections; lower panels: cross sections. (Hair is autofluorescent.) Scale bar = 40 μm and 50 μm in upper and lower panels, respectively. (c) A dramatic reduction in the number of nociceptor free nerve endings that penetrate the epidermis of hindlimb footpad glabrous skin of Hoxd1 −/− mice was detected by anti-CGRP staining, which labels peptidergic nociceptors. Arrows indicate epidermal nociceptor endings. Scale bar = 40 μm. (d) Double fluorescent labeling of Mrgprb4 mRNA and CGRP protein in L3 DRGs of WT and Hoxd1 −/− mice. (e) Quantification of the number (±SEM) of transverse lanceolate endings per mm 2 skin area in serial sections of hindlimb back thigh hairy skin. * p < 0.001 by Student’s t -test. (f) Percentage (±SEM) of Mrgprb4 + neurons in WT and Hoxd1 −/− mice that co-express CGRP or TrkA . * p < 0.001 by Student’s t -test. (g) Quantification of the percentage (±SEM) of lumbar DRG neurons that express Mrgprb4 . * p < 0.001 by Student’s t -test. (h) Quantification of the average (±SEM) number of CGRP + free nerve endings crossing the dermal-epidermal boundary per unit length (300 μm) of hindlimb footpad glabrous skin. * p < 0.005 by Student’s t -test.

Article Snippet: Primary antibodies used include: rabbit anti-mouse TrkA (Millipore, 1:1000), rabbit anti-chicken TrkA and TrkC (gifts from Dr. Frances Lefcort, Montana State University, 1:1000), rabbit (Millipore, 1:1000) or mouse anti-peripherin (Millipore, 1:500), rabbit anti-CGRP (Millipore, 1:1000), rabbit anti-PGP9.5 (Millipore, 1:1000), rabbit anti-Parvalbumin (Swant, 1:2000), rabbit anti-GFP (1:2000, Invitrogen), chicken anti-LacZ (Promega, 1:500), rabbit anti-Ret (Immuno-Biological Laboratories, 1:1000), mouse anti-chick Neurofilament 160kD (Zymed, 1:500) and rabbit anti-Neurofilament 200 (Millipore, 1:1000).

Techniques: Staining, Labeling

Journal: Nature neuroscience

Article Title: An evolving NGF–Hoxd1 signaling pathway mediates development of divergent neural circuits in vertebrates

doi: 10.1038/nn.2710

Figure Lengend Snippet: (a) DNA constructs used for in ovo electroporation. (b) Sox10 enhancer-driven in ovo electroporation directs ectopic gene expression in developing chicken DRGs but not in the spinal cord. Left panel: A whole-mount lateral view of GFP fluorescence in stage 26 chick embryos 3 days after electroporation. Right panel: Transverse sections of embryos processed as in the left panel, stained with anti-GFP. sp, spinal cord. drg, dorsal root ganglion. (c) The pattern of central projections of chick nociceptors is altered when Hoxd1 is expressed in the chick DRG. Upper panel: A spinal cord section labeled with anti-TrkA at stage 35 (e9); Lower panels: high-magnification views of the boxed areas of the same spinal cord section. The left side is control, and DRGs of the right side of the embryo are electroporated with Hoxd1 . Scale bar = 50 μm and 25 μm in low- and high-magnification panels, respectively. (d) Quantification of (c) shows the ratio between spinal cord areas occupied by TrkA + fibers in the electroporated side versus the control side. * p = 0.0001 by Student’s t -test (n > 4 embryos for each group; sections are 25 μm thick and are sampled at least 200 μm apart). cHoxd1 , chicken Hoxd1 gene. cΔ , a chicken Hoxd1 gene construct lacking the homeobox motif. mHoxd1 , mouse Hoxd1 gene. mΔ , a mouse Hoxd1 construct lacking homeobox. (e) Diagram of the different patterns of spinal cord innervation by mammalian and avian nociceptors.

Article Snippet: Primary antibodies used include: rabbit anti-mouse TrkA (Millipore, 1:1000), rabbit anti-chicken TrkA and TrkC (gifts from Dr. Frances Lefcort, Montana State University, 1:1000), rabbit (Millipore, 1:1000) or mouse anti-peripherin (Millipore, 1:500), rabbit anti-CGRP (Millipore, 1:1000), rabbit anti-PGP9.5 (Millipore, 1:1000), rabbit anti-Parvalbumin (Swant, 1:2000), rabbit anti-GFP (1:2000, Invitrogen), chicken anti-LacZ (Promega, 1:500), rabbit anti-Ret (Immuno-Biological Laboratories, 1:1000), mouse anti-chick Neurofilament 160kD (Zymed, 1:500) and rabbit anti-Neurofilament 200 (Millipore, 1:1000).

Techniques: Construct, In Ovo, Electroporation, Gene Expression, Fluorescence, Staining, Labeling, Control