Notes: This paper describes an assay to test BDNF dissociation from FluoSpheres (amine-modified microspheres, 1μM diameter; Molecular Probes). BDNF was covalently attached to the microspheres. Release of BDNF was tested in supernatants from the BDNF-microspheres in PBS, in complete media, and in culture with rat Dorsal root ganglia. Promega’s BDNF Emax® ImmunoAssay System was used to determine the amounts of BDNF in these samples. The DeadEnd™ Fluorometric TUNEL System was used to examine rat Dorsal root ganglia cells that had been transfected with either dynamitin or β-Galactosidase. In the TUNEL experiments, cells were fixed in 4% paraformaldehyde and counterstained with DAPI. Cells were also stained with Promega’s Anti-β-Galactosidase, Purified Monoclonal Antibody. For these immunocytochemical stains, a 1:500 dilution of Anti-β-Galactosidase antibody and a 1:1,500 dilution of a secondary antibody conjugated to Alexa Fluor-488 (Molecular Probes) were used. The stains were visualized by fluorescence microscopy. (3055)

Notes: Anti-β-Galactosidase Monoclonal Antibody was used to immunohistochemically stain stomach sections from C57BL/6 mice transplanted with ROSA26 marrow and infected with Helicobacter pylori. The Anti-β-Galactosidase antibody preferentially stained H. pylori β-galactosidase, helping to demonstrate infection of the mouse stomachs by H. pylori. (3413)

Notes: In this study, the authors used the Anti-β-Galactosidase mAb to distinguish modified embryonic stem cells from wild-type host cells in mouse chimeras. During the creation of the chimeras, β-galactosidase was used as a marker for genetically modified stem cells.  In these experiments whole embryos were fixed in 4% paraformaldehyde with 30% sucrose and mounted with OCT medium. The Anti-β-Galactosidase mAb was used at a dilution of 1:1000.  (3199)

Notes: Primary dissociated mouse embryonic cortex E14 cells or cultured astrocytes were fixed with paraformaldehyde and stained with the anti β-galactosidase mAb in immunocytochemistry studies.  The antibody was used at a 1:2000 dilution. (2581)

Notes: The β-galactosidase mAb was used to stain Drosophila wings that contained cells expressing a Gal 4-Frizzled construct. (2583)

Notes: The authors of this study identified pi cells based on the specific expression of a lin-11::lac-Z transgene, detected using the Anti-β-Galactosidase mAb. In these experiments, whole C. elegans worms were mounted on slides with antifade (Molecular Probes) and counterstained with DAPI. (3198)

Notes: The authors seek to determine the developmental roles played by differential expression of different β-tubulin proteins during embryogenesis in Drosophila. Fly strains with a β3-tubulin lacZ enhancer trap were used to map the β3 tubulin gene and to express β-galactosidase in a pattern similar to β3 tubulin. Expression of β-galactosidase expression was localized by immunohistochemistry with the Anti-β-Galactosidase mAb on embryos fixed in 4% paraformaldehyde. (2445)

Notes: The authors use a T7 polymerase-driven full-length hepatitis C viral cDNA plasmid to reproduce the early steps of the HCV life cycle in vivo. The positive control plasmid used in these experiments contained a T7 promoter flanking the β-galactosidase gene. Expression of HCV proteins and the control β-galactosidase protein in CV-1 and HepG2 cells was detected by Western blot analysis. The Anti-β-Galactosidase mAb was used at a 1:5,000 dilution. (2417)

Notes: The RPN4 protein is involved in the regulation of levels of proteasomal subunits in Saccharomyces cerevisiae. The protein was found to be short-lived by fusing the RPN4 degradation signal to the long-lived β-galactosidase protein. Levels of this fusion protein were quantitated by immunoprecipitation of equal amounts of TCA-insoluble 35S labeled protein using the Anti-β-Galactosidase mAb. Yeast cells were lysed by vortexing with glass beads in lysis buffer (1% Triton® X-100/0.15 M NaCl/1 mM EDTA/50 mM Na-Hepes, pH 7.5) containing protease inhibitors prior to immunoprecipitation. (2446)

Notes: Two potential nuclear localization signals in the adenomatous polyposis coli  gene were scrutinized by inserting these signals into a β-galactosidase expression vector. Mouse L cells and Cos7 cells were transfected with the β-galactosidase construct and immunostained to monitor nuclear transport. The Anti-β-Galactosidase mAb (1:1,000 dilution) and a goat anti-mouse IgG-FITC were used to immunostain the transfected cells. (2423)

Notes: The Serine-Threonine Phosphatase Assay System was used to monitor dephosphorylation of two phosphopeptides derived from the SCR homeodomain. Dephosphorylation was performed using the catalytic subunit of the PP2A protein. Authors also used pSI for cloning, protein kinase A enzyme, PKA and PKC assays using biotinylated peptides (SignaTECT® Protein Kinase assay systems), anti-β-galactosidase for Drosophila embryo staining, and TNT® Quick Coupled Transcription/Translation systems to produce mutant and wild type protein for gel shifts. (2150)

Notes: LIM1 is a homeodomain protein involved in leg development in Drosophila melanogaster. Fly strains with a Lim1-lacZ enhancer trap were stained by immunohistochemistry for β-galactosidase to localize LIM1 expression. Immunohistochemistry on intact flies was performed using Promega's Anti-β-Galactosidase mAb. (2444)

Notes: Knockout mice deficient in the tyrosine kinase receptor Kit, which is critical for the development of melanocytes from neural crest-derived precursor cells, were generated. The Kit encoding region was replaced in these mice with the LacZ gene, a convenient marker for Kit expression. Kit-LacZ homozygous mice were identified by PCR using Promega's Taq DNA Polymerase to amplify a 800bp band corresponding to LacZ and to confirm the absence of the 148bp band of Kit. Primary neural crest cell cultures from knockout mouse embryos were immunostained with the Anti-β-Galactosidase mAb to localize Kit expression. Cells were fixed in 4% paraformaldehyde and permeabilized with 0.1% Triton®-X-100 prior to immunostaining. (2422)

Notes: The Anti-β-Galactosidase mAb was used in indirect immunofluorescence experiments. (0272)

Notes: The 2.5S Nerve Growth Factor was used to culture rat dorsal root ganglia neurons. The primary rat cells were infected with an adenovirus expressing E. coli β-galactosidase (lacZ) and the enzyme was detected by immunocytochemistry with the Anti-β-Galactosidase mAb. Primary cells infected with either an HSP27 adenovirus or LacZ adenovirus were withdrawn from NGF and survival assayed 48 hours later with the CellTiter 96^® Non-Radioactive Cell Proliferation Assay. Only those cells expressing HSP27 showed some survival and percent survival was dependent upon the multiplicity of infection. (0788)

Notes: The Anti-β-Galactosidase mAb was used for immunostaining of 12µm thick free-floating rat brain sections, fixed with paraformaldehyde by a referenced method. The antibody was used at a 1:1000 dilution. The LacZ was introduced by an adenoviral vector. (0122)

Notes: Regulation of glutamyl-tRNA reductase (HemA), which catalyzes the first committed step in heme synthesis appears to be through stabilization of the protein rather than transcriptional regulation. Western blot analysis of HemA and a HemA-LacZ fusion protein revealed that the HemA protein accumulates under low heme conditions. The Anti-B-Galactosidase mAb was used to detect the HemA-LacZ fusion protein in Salmonella typhimurium and Escherichia coli. The same antibody was also used for immunoprecipitation of the HemA-LacZ fusion protein. (2420)

Notes: BHK-21 cells were transfected with either p21-activated kinase expression vectors or control vectors expressing LacZ. LacZ expression was detected with Anti-β Galactosidase mAb. A lot of detail is provided for cell fixation, permeabilization and antibody incubation for the immunocytochemistry. (0435)

Notes: The authors examine the colocalization of several RNA-binding proteins and poly(A)(+) RNA at mammalian stress granules, cytoplasmic foci that result from an environmental stress. The effects of wild-type and mutant eIF-2 on stress granule formation was also examined. COS cells were transiently transfected with a ß-galactosidase reporter plasmid and plasmids encoding wild-type and mutant eIF-2. Cell lysates were subjected to Western blot analysis with the Anti–ß-Galactosidase mAb (1:2000 dilution) to monitor ß-galactosidase expression. (2419)

Notes: The Anti β-Galactosidase mAb was used in immunostaining of fly's eyes. (0873)

Notes: The putative pheromone receptor gene VRi2 was replaced with a lacZ expression cassette in transgenic mice to localize expression of the receptor in the vomeronasal system, which mediates pheromonal effects in mammals. Immunohistochemistry of the mouse accessory olfactory bulb was performed with the Anti-β-Galactosidase mAb at a concentration of 7 µg/ml. Tissues were fixed in 2% formaldehyde and cut into 7 micrometer sections prior to staining. (2443)

Notes: Promega's Anti β-Galactosidase monoclonal antibody was used to detect lacZ expression in situ using whole mount Drosophila embryos. The conditions used for mounting, fixing, and probing with the antibodies used is given in the Materials and Methods. The Anti β-Galactosidase mAb is used at a dilution of 1:1000. (2169)

Notes: In an enhancer-trap experiment, the Anti β-Galactosidase mAb was used for immunostaining to identify mutants that carry insertions of a β-Galactosidase reporter gene near an enhancer element that enhances the expression in the ring gland of Drosophila. T4 DNA ligase, Taq DNA Polymerase and Proteinase K were also used in the studies. (1067)

Notes: Anti-β-Galactosidase mAb was used for immunohistochemistry (IHC) staining of Drosophila wing discs 1 hour after puparium formation (APF). (0295)

Notes: Adult rat hippocampus-derived neural progenitor (AHPC) cells, engineered to express the marker B-galactosidase, were grafted into the vitreous cavity of rat eyes to determine the cells' ability to integrate into the retina. AHPC were detected in rat retinal tissues by immunohistochemistry. Eyes grafted with AHPC cells were fixed with 4% paraformaldehyde and permeabilized with 0.3% Triton® X-100; 20µm sections were stained with the Anti-B-Galactosidase mAb (1:5000 dilution). (2424)