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Vectasheld

Manufactured by Vector Laboratories
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Sourced in United States

Vectasheld is a microscope slide mounting medium designed for use in immunohistochemistry and other microscopy applications. It is a water-based, non-fluorescent medium that provides excellent clarity and preservation of tissue samples.

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Lab products found in correlation

Te2000 u, by Nikon (1 mentions) In situ apoptosis detection kit, by Takara Bio (1 mentions) 4 6 diamidino 2 phenylindole (dapi), by Lonza (1 mentions) Bz 8100, by Keyence (1 mentions)

3 protocols using vectasheld

1

Apoptosis Detection by TUNEL Assay

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The apoptotic signal was determined by TUNEL assay using a commercially available kit (In Situ Cell Death Detection Kit, fluorescein, Roche, Indianapolis, IN, USA), following manufacturer recommendations. Briefly, the fixed cells on the slides were washed three times for 5 min with PBS- and permeabilized with 0.1% (v/v) Triton X-100 containing 0.1% (w/v) sodium citrate for 2 min on ice. Samples were then incubated in 50 μL of TUNEL reaction mixture for 1 h at 37 °C in a dark and humidified atmosphere. Slides were washed three times with PBS- and cover slips were mounted using mounting medium (VECTASHELD®, Vector Laboratories, and Burlingame, CA, USA). Positive TUNEL GFP staining was observed under a fluorescence microscope (TE2000U, Nikon, Tokyo, Japan) using the B-2A filter (450–490 nm excitation filter, 505 nm dichroic mirror, 520 nm band pass filter).
El Andaloussi A., Igboeli P., Amer A, & Al-Hendy A. (2018). Intravenous Infusion of Nucleated Peripheral Blood Cells Restores Fertility in Mice with Chemotherapy-Induced Premature Ovarian Failure. Biomedicines, 6(3), 93.
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2

Immunohistochemical Characterization of Prostate Cancer

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Primary prostate cancer tissues were obtained anonymously as described previously (1 (link)) and from the Molecular Pathology Shared Resource of the Herbert Irving Comprehensive Cancer Center. Tissues were used by IRB approval. Tissues were not selected in any way other than the diagnosis of prostate cancer that had been treated by prostatectomy. Deparaffinized slides were pretreated in 10 mM citrate buffer pH 6.0 in a steamer for 40 min. The cells were permeabilized with 0.5% Triton X-100 for 15 min and then 0.3% H2O2 in PBS for 60 min to quench endogenous peroxidase activity. The slides were incubated with rabbit NKX3.1 (1:2000) antibody and murine histone H1 (1:1000) overnight, rinsed with PBS+ 5% Tween 20, then incubated with biotinylated-anti-mouse IgG antibody (1:200) for 30 min. After three rinses with PBS + 5% Tween 20 + 1% goat serum for 5 min each, the slides were incubated with Texas-read-avidin (1:200) and secondary anti-rabbit IgG-horseradish peroxidase (1:200) antibody for 30 min. Fluorescence-plus-Tyramide (PerkinElmer) was applied on slides for 6 min, and ToPro3 (1:1000) for 5 min. Cover slips were mounted with Vectasheld (Vector).
Bowen C., Zheng T, & Gelmann E.P. (2015). NKX3.1 Suppresses TMPRSS2-ERG Gene Rearrangement and Mediates Repair of Androgen Receptor-Induced DNA Damage. Cancer research, 75(13), 2686-2698.
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3

TUNEL Assay for Apoptosis Detection

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To investigate the possible occurrence of apoptosis, a terminal deoxynucleotidyl transferase mediated dUTP-biotin nick end labeling (TUNEL) assay [49] (link) was performed using an in situ apoptosis detection kit (Takara, Kyoto, Japan). All samples were digested with a proteinase K solution (10 µg/ml proteinase K in PBS) for 15 min at room temperature to increase their permeability. After being washed twice with PBS for 5 min, these sections were incubated for 1 h with terminal deoxynucleotidyl transferase and fluorescein isothiocyanate-labeled dUTPs in a humidified chamber at 37°C to label the exposed 3′-hydroxyl ends of fragmented nuclear DNA. After terminating the reaction, the sections were again washed twice with PBS for 5 min. Sections were counterstained with 4, 6-Diamidino-2-phenylindole (DAPI; 30 nM solution in PBT) (Lonza, Walkersville, MD, USA) in the dark for 30 min at room temperature. The stained sections were mounted in Vectasheld (Vector Laboratories, Burlingame, CA, USA) and examined using a fluorescent light microscope (BZ-8100 Keyence, Osaka, Japan).
Niitsu S., Toga K., Tomizuka S., Maekawa K., Machida R, & Kamito T. (2014). Ecdysteroid-Induced Programmed Cell Death Is Essential for Sex-Specific Wing Degeneration of the Wingless-Female Winter Moth. PLoS ONE, 9(2), e89435.
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