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Neg50 mounting medium

Manufactured by Thermo Fisher Scientific
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The NEG50 mounting medium is a water-soluble, glycol-based liquid used for mounting samples on microscope slides. It is formulated to provide a clear, transparent medium for preserving and protecting specimens during microscopic examination.

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Lab products found in correlation

Cryojane taping system, by Leica Biosystems (2 mentions) Avertin, by Merck Group (1 mentions) Ab21176, by Abcam (1 mentions) Cryojane taping system, by Leica camera (1 mentions) Cellsens dimension imaging, by Olympus (1 mentions) Rat anti endomucin, by Thermo Fisher Scientific (1 mentions) Alexa fluor 488 or 594 conjugated secondary antibodies, by Jackson ImmunoResearch (1 mentions) Rat anti pdgfrβ, by Thermo Fisher Scientific (1 mentions) Alexa fluor 594 conjugated secondary antibody, by Jackson ImmunoResearch (1 mentions) Mab1398z, by Merck Group (1 mentions) Alexa fluor 488, by Jackson ImmunoResearch (1 mentions)

4 protocols using neg50 mounting medium

1

Viral Particle Delivery and Tissue Fixation

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Mice were injected with 1 × 1011 or 6 × 1010 particles of virus in 200 μl of saline via the tail vein. At designated times post-vector injection, mice were induced into deep anesthesia with 2.5% 2,2,2-tribromoethanol (Avertin; Sigma-Aldrich, St. Louis, MO), and the thorax was opened. A 24-gauge round ball-tipped stainless oral gavage needle was inserted into the left ventricle, and the mouse was perfused with 30 ml of 10% PBS-buffered formalin. Organs were removed and cut into 1- to 2-mm-thick slices. Organ slices underwent postperfusion fixation in formalin at room temperature for 1 to 2 h. Harvested organ tissues were prepared in dual sets of frozen and paraffin embedded sections. For frozen sections, organ slices were cryopreserved in 30% sucrose in PBS at 4°C overnight, embedded in NEG50 mounting medium (Thermo Fisher Scientific, Waltham, MA), and frozen in a 2-methylbutane-containing glass beaker prechilled in liquid nitrogen. Fixed tissues were also switched to graded alcohol and xylene series and embedded in paraffin.
Lu Z.H., Dmitriev I.P., Brough D.E., Kashentseva E.A., Li J, & Curiel D.T. (2020). A New Gorilla Adenoviral Vector with Natural Lung Tropism Avoids Liver Toxicity and Is Amenable to Capsid Engineering and Vector Retargeting. Journal of Virology, 94(10), e00265-20.
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2

Immunohistochemical Analysis of Luciferase in Frozen Mouse Tissues

Cited 2 times
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Harvested mouse organs were fixed with 10% formalin phosphate solution and preserved in 30% sucrose in PBS at 4 °C overnight. The fixed tissues were embedded in NEG50 mounting medium (Thermo Scientific, Waltham, MA USA) then frozen in liquid nitrogen pre-chilled 2-methylbutane. Tissue slide sectioning was performed from frozen tissues using the CryoJane taping system (Leica CM1900). All tissues slides were subjected to IHC analysis with specific primary antibodies (rabbit anti- luciferase [1:200, catalog number ab21176; Abcam, Boston, MA USA]) and using Alexa Fluor 488- or Alexa Fluor 594-conjugated secondary antibodies (1:400, #103-545-155 or #711-585-152; Jackson ImmunoResearch Laboratories, West Grove, PA USA). Images were analyzed using CellSens Dimension imaging software (Olympus Soft Imaging Solutions). All processes were in accordance with previously published protocols [42 (link),45 (link)].
Lee M., Rice-Boucher P.J., Collins L.T., Wagner E., Aulisa L., Hughes J, & Curiel D.T. (2022). A Novel Piggyback Strategy for mRNA Delivery Exploiting Adenovirus Entry Biology. Viruses, 14(10), 2169.
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3

Targeted Adenoviral Delivery and Tissue Analysis

Cited 1 time
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Mice were administrated with 1×10 11 VP of triple targeted Ad via tail-vein injection. Three days post virus infection, mice were anesthetized and tissues (Liver, Lung, Pancreas, Spleen, Kidney, Small Bowel, Heart, Muscle and Brain) harvested for immunofluorescence staining. For frozen sections, organ slices were cryo-preserved in 30% sucrose in PBS at 4oC overnight, embedded in NEG50 mounting medium (Thermo Fisher Scientific), and then frozen in a liquid nitrogen pre-chilled 2-methylbutane containing bucket. Sectioning of frozen organs was carried out using the CryoJane taping system (Leica Biosystems Inc). All frozen section slides were subject to immunofluorescence staining analysis in accordance with the standard techniques (8 (link)). Primary antibodies used in this study included hamster anti-CD31 (EMD Millipore), rat anti-endomucin (eBioscience), rat anti-PDGFRβ (eBioscience), rabbit anti-HSVtk (Dr. Summers’s lab) and Alexa Fluor 488 or 594-conjugated secondary antibodies (Jackson ImmunoResearch Laboratories).
Lee M., Lu Z.H., Li J., Kashentseva E.A., Dmitriev I.P., Mendonca S.A, & Curiel D.T. (2020). Targeting tumor neoangiogenesis via targeted adenoviral vector to achieve effective cancer gene therapy for disseminated neoplastic disease. Molecular cancer therapeutics, 19(3), 966-971.
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4

Immunostaining of Mouse Tissue Samples

Cited 1 time
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Mice were anesthetized and the tissues harvested (liver, spleen, small intestine and omentum [tumor cells]) for immunofluorescence (IF) or immunohistochemistry (IHC) staining. Organ slices were cryo-preserved in 30% sucrose in PBS at 4 °C overnight for frozen tissues, embedded in NEG50 mounting medium (Thermo Fisher Scientific). The tissues were then frozen in a liquid nitrogen pre-chilled 2-methylbutane. Sectioning of frozen tissues was carried out using the CryoJane taping system (Leica Biosystems Inc). All tissues slides were subject to IHC analysis with specific primary Abs (Armenian hamster anti-CD31 [1:1000; catalog number MAB1398Z; MilliporeSigma], rat anti-endomucin [1:1000; catalog number 14-5851-81; eBioscience], human anti-B7-H3 (CD276) [1:200, catalog number AF1027; R&D Systems antibody], rabbit anti-firefly luciferase [1:200, catalog number ab21176; Abchem], and used the Alexa Fluor 488- or Alexa Fluor 594-conjugated secondary antibodies (Jackson ImmunoResearch Laboratories, West Grove, PA) for images acquisition, in accordance with the standard protocols as published previously [7 (link),30 ]. Images were analyzed by CellSens Dimension imaging software (Olympus Soft Imaging Solutions).
Frischer M.E., Danforth J.M., Healy M.A, & Saunders F.M. (2000). Whole-Cell versus Total RNA Extraction for Analysis of Microbial Community Structure with 16S rRNA-Targeted Oligonucleotide Probes in Salt Marsh Sediments. Applied and Environmental Microbiology, 66(7), 3037-3043.
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