Journal:

Article Title: Enkephalins, dynorphins and ?-endorphin in the rat dorsal horn: an immunofluorescence colocalization study

doi: 10.1002/cne.22130

Figure Lengend Snippet: The β-endorphin antibody [Mu(-5)N] was used to stain sections through the arcuate nucleus (ArcN) of wild type mice (WT, A) and β-endorphin deficient mice (β-endorphin KO, B). Note the complete disappearance of β-endorphin immunoreactivity in the β-endorphin defiant mice. Images were taken using a 20x objective and a digital camera.

Article Snippet: The species, sources, dilutions and immunogen of the primary antibodies are indicated in . table ft1 table-wrap mode="anchored" t5 caption a7 antibody species source catalog number dilution immunogen β-endorphin rabbit RIKEN Mu(-5)N 1:1000 β-endorphin minus YGGF N-terminus (CTSEKSQTPLVTLFKNAIIKNAHKKGQ) CGRP(g) goat Santa Cruz sc-8856 1:5000 human CGRP, N-terminus (ACDTATCVTHRLAGLLSRSG) CGRP(m) mouse * CURE 4901 1:10000 rat α-CGRP (SCNTATCVTHRLAGLLSR SGGVVKDNFVPTNVGSEAF-NH 2 ) CGRP(r) rabbit Dr. Sternini -- 1:10000 rat CGRP 23-37 (VKDNFVPTNVGSEAF) dynorphin A rabbit Peninsula IHC8730 1:200 pig dynorphin A 1-17 (YGGFLRRIRPKLKWDNQ) enkephalin mouse * Millipore MAB135 1:1000 YGGFL enkephalin, FMR- rabbit CSORDA -- 1:5000 YGGFMR MOR rabbit ImmunoStar 24216 1:6000 MOR-1 residues 384-398 (NHQLENLEAETAPLP) NK1R rabbit CURE 94168 1:3000 NK1R residues 393-407 (KTMTESSSFYSNMLA) pan-opioid (3-E7) mouse * Gramsch 3-E7 1:1000 YGGFM PKCγ rabbit Santa Cruz sc-211 1:1000 mouse PKCγ residues 679-697 (DFVHPDARSPTSPVPVPVM) prodynorphin guinea pig Neuromics GP10110 1:2000 rat pro-dynorphin residues 235-248 (SQENPNTYSEDLDV) SP(g) goat Santa Cruz sc-9758 1:2000 substance P (RPKPQQFFGLM-NH 2 ) SP(r) rabbit CURE 3801 1:1000 substance P (RPKPQQFFGLM-NH 2 ) VGLUT2 guinea pig Millipore AB5907 1:2000 rat VGLUT2 residues 565-582 Open in a separate window * monoclonal antibody Antibody sources: CURE: Digestive Diseases Research Center (CURE), UCLA, Los Angeles, CA; Center for the Study of Opioid Receptors and Drugs of Addiction (CSORDA), UCLA, Los Angeles, CA; Gramsch Laboratories, Schwabhausen, Germany; ImmunoStar, Hudson, WI; Millipore, Billerica, MA; RIKEN Brain Science Institute, Wako, Saitama, Japan; Neuromics, Edina, MN; Peninsula Laboratories Inc. (now Bachem), San Carlos, CA; Santa Cruz Biotechnology, Santa Cruz, CA; Dr. Catia Sternini, Department of Medicine, UCLA, Los Angeles, CA.

Techniques: Staining

Journal:

Article Title: Enkephalins, dynorphins and ?-endorphin in the rat dorsal horn: an immunofluorescence colocalization study

doi: 10.1002/cne.22130

Figure Lengend Snippet: Coronal sections from the lumbar spinal cord (L5-L6) were labeled with antibodies against β-endorphin [Mu(-5)N] and VGLUT2. Only the β-endorphin staining is shown. The VGLUT2 label was used to define the edge of the gray matter, indicated by the lines. Confocal sections were obtained with a 20x objective at 1.18 μm intervals, and were de-blurred using blind deconvolution. Dorsal side is up. A, dorsal horn (10 sections); a rare β-endorphin fiber is shown. B, lamina X (9 sections), β-endorphin immunoreactive puncta are present around the central canal. C, ventral horn (10 sections); some β-endorphin immunoreactive puncta and fibers are observed in the grey and the white matter; large cell bodies were stained at lower intensity. Scale bars are 50 μm. See Table 2 for abbreviations.

Article Snippet: The species, sources, dilutions and immunogen of the primary antibodies are indicated in . table ft1 table-wrap mode="anchored" t5 caption a7 antibody species source catalog number dilution immunogen β-endorphin rabbit RIKEN Mu(-5)N 1:1000 β-endorphin minus YGGF N-terminus (CTSEKSQTPLVTLFKNAIIKNAHKKGQ) CGRP(g) goat Santa Cruz sc-8856 1:5000 human CGRP, N-terminus (ACDTATCVTHRLAGLLSRSG) CGRP(m) mouse * CURE 4901 1:10000 rat α-CGRP (SCNTATCVTHRLAGLLSR SGGVVKDNFVPTNVGSEAF-NH 2 ) CGRP(r) rabbit Dr. Sternini -- 1:10000 rat CGRP 23-37 (VKDNFVPTNVGSEAF) dynorphin A rabbit Peninsula IHC8730 1:200 pig dynorphin A 1-17 (YGGFLRRIRPKLKWDNQ) enkephalin mouse * Millipore MAB135 1:1000 YGGFL enkephalin, FMR- rabbit CSORDA -- 1:5000 YGGFMR MOR rabbit ImmunoStar 24216 1:6000 MOR-1 residues 384-398 (NHQLENLEAETAPLP) NK1R rabbit CURE 94168 1:3000 NK1R residues 393-407 (KTMTESSSFYSNMLA) pan-opioid (3-E7) mouse * Gramsch 3-E7 1:1000 YGGFM PKCγ rabbit Santa Cruz sc-211 1:1000 mouse PKCγ residues 679-697 (DFVHPDARSPTSPVPVPVM) prodynorphin guinea pig Neuromics GP10110 1:2000 rat pro-dynorphin residues 235-248 (SQENPNTYSEDLDV) SP(g) goat Santa Cruz sc-9758 1:2000 substance P (RPKPQQFFGLM-NH 2 ) SP(r) rabbit CURE 3801 1:1000 substance P (RPKPQQFFGLM-NH 2 ) VGLUT2 guinea pig Millipore AB5907 1:2000 rat VGLUT2 residues 565-582 Open in a separate window * monoclonal antibody Antibody sources: CURE: Digestive Diseases Research Center (CURE), UCLA, Los Angeles, CA; Center for the Study of Opioid Receptors and Drugs of Addiction (CSORDA), UCLA, Los Angeles, CA; Gramsch Laboratories, Schwabhausen, Germany; ImmunoStar, Hudson, WI; Millipore, Billerica, MA; RIKEN Brain Science Institute, Wako, Saitama, Japan; Neuromics, Edina, MN; Peninsula Laboratories Inc. (now Bachem), San Carlos, CA; Santa Cruz Biotechnology, Santa Cruz, CA; Dr. Catia Sternini, Department of Medicine, UCLA, Los Angeles, CA.

Techniques: Labeling, Staining

Journal: Free radical biology & medicine

Article Title: Blue light (λ=453 nm) nitric oxide dependently induces β-endorphin production of human skin keratinocytes in-vitro and increases systemic β-endorphin levels in humans in-vivo.

doi: 10.1016/j.freeradbiomed.2019.09.022

Figure Lengend Snippet: Fig. 2. Impact of blue light irradiation on p53, pro-opiomelanocortin (POMC), pro-hormone convertase 1/3 (PC1/3) as well as pro-hormone convertase 2 (PC2) protein expression of human dermal keratinocytes. Shown are values of relative protein concentration (related to the untreated control) of human skin keratinocyte cultures exposed to 20 (white bars), 40 (gray bars) or 80 J/cm2 (black bars) of blue light with a maximum intensity at 453 nm. P53 (A), POMC (B), PC1/3 (C) as well as PC2 protein (D) levels were assessed by Western blot analysis and normalized to GAPDH expression. Bars represent the mean ± SEM of 10–12 individual experiments.

Article Snippet: In order to detect POMC, PC 1/3, PC 2 and p53 we used Goat Polyclonal Anti-POMC Antibody (Novus Biologicals), Rabbit Polyclonal Anti-PC 1/ 3 Antibody (Cell Signaling), Rabbit Monoclonal Anti-PC 2 Antibody (Cell Signaling) and Mouse Monoclonal Anti-p53 Antibody (Thermo Scientific).

Techniques: Irradiation, Expressing, Protein Concentration, Control, Western Blot

Journal: Brain structure & function

Article Title: Neuronal connections of the central amygdalar nucleus with refeeding-activated brain areas in rats

doi: 10.1007/s00429-017-1501-4

Figure Lengend Snippet: Antibody Characterization

Article Snippet: Because PHA-L and CTB are not present in the brain enogenously, the specificities of PHA-L and CTB antisera were verified by the lack of any labeling in brain sections from animals that were not injected with PHA-L and CTB. table ft1 table-wrap mode="anchored" t5 Name of the antibody Structure of the immunogen Manufacturing Dilution c-Fos Synthetic peptide (SGFNADYEASSS RC) corresponding to amino acids 4–17 of human c-Fos Merck Millipore, rabbit, polyclonal, #cat. PC38 RRID AB_213663 1:10000 CTB Purified CTB isolated from Vibrio cholerae List Biological Laboratories, goat, polyclonal, #cat. 703 RRID AB_2314252 1:5000 PHAL Purified PHAL Vector Laboratories, goat, polyclonal, #cat. AS-2224 RRID AB_2315141 1:5000 POMC Amino acids of 27–52 of porcine POMC precursor Phoenix Pharmaceuticals, rabbit, polyclonal, #cat. H-029-30 RRID AB_2307442 1:2000 HuCD Recognizes human HuC/HuD neuronal protein and the expressed product of the ELAVL3 gene Molecular Probes, mouse, monoclonal, #cat. A-21271 RRID AB_221448 1:500 CRH human/rat (h/r)CRF peptide (SEEPPI SLDLTFHLLREVL EMARAEQLAQQA HSNRKLMEII-NH2), Peninsula Laboratories International, Inc, rabbit, polyclonal, #cat. T-4037 RRID AB_2314240 1:3000 RFP The immunogen is a Red Fluorescent Protein (RFP) fusion protein corresponding to the full length amino acid sequence (234aa) derived from the mushroom polyp coral Discosoma.

Techniques: Purification, Isolation, Plasmid Preparation, Sequencing, Derivative Assay

Journal: Cureus

Article Title: Effects of Pain Relief Through Minimal Exercise Intervention in a Rat Model of Neuropathic Pain

doi: 10.7759/cureus.62897

Figure Lengend Snippet: The graph shows β-endorphin/met-enkephalin positive cell area ratio (E), with a significant increase in the Ex group compared to the No-Ex group. Data are expressed as mean ± SE. * p < 0.05, ** p < 0.01. Scale bar = 50 μm (all panels). Aq: aqueduct; Ex: exercise; DAB: diaminobenzidine

Article Snippet: The sections were rinsed three times for 5 min each in PBS and incubated individually at 4°C with one of the following antibodies: rabbit anti-ionized calcium-binding adaptor molecule 1 (IBA1) antibody (1:2000, rabbit polyclonal, Wako Pure Chemical Industries, Osaka, Japan), a marker for microglia; rabbit anti-glial fibrillary acidic protein (GFAP) antibody (1:1000, rabbit polyclonal, Shima Research Institute, Tokyo, Japan), a marker for astrocytes; goat anti-CCR2 antibody (1:1500, goat polyclonal (1:600, mouse monoclonal, Proteintech Group, Inc., USA), a marker for TRAF6; anti-BDNF antibody, a marker for cranial nerve-derived neurotrophic factor (BDNF) (1:600, mouse monoclonal, Proteintech Group, Inc., USA); and anti-BDNF antibody (1:200, rabbit polyclonal, Bioss Antibodies Inc.); and anti-β-endorphin/met-enkephalin antibody, a marker for endogenous opioids (1:50; Santa Cruz Biotechnology, Inc.).

Techniques: