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VECTASHIELD® HardSet™ Antifade Mounting Medium with DAPI

H-1500

Still using VectaShield® HardSet™?

Try our upgraded product VECTASHIELD® Vibrance™ Antifade Mounting Medium

SKU Unit Size Price Qty
H-1500-10 10 ml
$170.00

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Description

VECTASHIELD HardSet Antifade Mounting Medium with DAPI is a unique, stable formula for preserving fluorescence. This medium prevents rapid photobleaching of fluorescent proteins and fluorescent dyes. VECTASHIELD HardSet Mounting Medium with DAPI hardens after coverslipping. After approximately 15 minutes at room temperature, the coverslip will become immobilized, and optimal antifade ability and refractive index will be achieved. 

Features:

  • Inhibits photobleaching of fluorescent dyes and fluorescent proteins
  • Ideal refractive index (1.46)
  • Ready-to-use - from the refrigerator to the benchtop
  • No warming necessary
  • Can be stored without sealing for long term analysis
  • Hardening formulation
  • DAPI counterstain

Specifications

More Information
Unit Size 10 ml
Applications Immunofluorescence, In situ hybridization, Cellular Imaging
Mounting Aqueous (Hardening)
Antifade Yes
Counterstain DAPI (blue fluorescence)
Cellular Stain Nucleus

Documents

Product FAQs

I am using VECTASHIELD® HardSet™ Mounting Medium with DAPI (Cat No. H-1500). The sections look good initially, however I do see bubble or vacuole formation occur under the coverslip after some time. Do you have any suggestions on how this situation can be remedied?

Fixed, thin cut (<10 um), glass mounted sections are recommended to be used in combination with this medium. Unfixed material, thicker sections or the use of gasket or chamber slides or well formats may increase the incidence of bubble formation when applying H-1500. If using thin sections as described, ensure an adequate volume of the medium is being applied to spread out under the coverslip. Storage of slides in the freezer (-20 °C) may also reduce the incidence of bubbles. For existing mounted sections with bubble formation, coverslips can be removed by soaking the slides in buffer, and then remounted using fresh media.

Does VECTASHIELD contain glycerol?

All of the VECTASHIELD formats we currently offer which includes regular non-setting VECTASHIELD, HardSet and Vibrance formulations each contain a percentage of glycerol.

Do I need to dehydrate the tissue sections before applying VECTASHIELD?

No form of tissue dehydration (e.g. air drying or ethanol exposure) is required nor recommended when applying VECTASHIELD. From our experience, the most optimal antifade actions of VECTASHIELD are obtained when the preparation is removed from the final buffer/water rinse, kept slightly wet/moist and then coverslipped with a small volume (25-50 uL) of VECTASHIELD.

Do I need to seal the perimeter of the coverslip after I apply VECTASHIELD?

That depends upon how long you wish to retain the slides and which VECTASHIELD formulation you are using. If you are using one of our non-setting VECTASHIELD products such as H-1000 or H-1200, then we suggest sealing the coverslip with plastic sealant or nail polish if you intend to keep the slides beyond a day or so. If you are using one of our setting/curing formulations such as VECTASHIELD HardSet or Vibrance, then in most cases when using thin cut (<10 um) tissue sections or cell monolayers, no sealing of the coverslip is requuired.

Related Products

Human Uterine section (FFPE): Stained for Desmin (green) and Cytokeratin (red) using VectaFluor™ Duet Double Labeling Kit (DK-8828), and vasculature using DyLight® 649 UEA I lectin (purple). Mounted in VECTASHIELD® Vibrance™ Antifade Mounting Medium. VECTASHIELD® Vibrance™ Antifade Mounting Medium Catalog #: H-1700
Human Uterine section (FFPE): Stained for Desmin (green) and Cytokeratin (red) using VectaFluor™ Duet Double Labeling Kit (DK-8828), and vasculature using DyLight® 649 UEA I lectin (purple). Mounted in VECTASHIELD® Vibrance™ with DAPI (blue) VECTASHIELD® Vibrance™ Antifade Mounting Medium with DAPI Catalog #: H-1800
Human Colon: Rabbit Anti-Cytokeratin (AE1/AE3) and Mouse Anti-Desmin detected simultaneously with VectaFluor Duet Double Labeling Kit (DK-8828); Vasculature detected using Dylight 649 UEA I lectin (purple). Mounted in VECTASHIELD PLUS. VECTASHIELD® PLUS Antifade Mounting Medium Catalog #: H-1900
Human Colon: Rabbit Anti-Cytokeratin (AE1/AE3) and Mouse Anti-Desmin detected simultaneously with VectaFluor Duet Double Labeling Kit (DK-8828); Dylight 488 (green)/Dylight 594 (red). Mounted in VECTASHIELD PLUS with DAPI (nuclei, blue). VECTASHIELD® PLUS Antifade Mounting Medium with DAPI Catalog #: H-2000
3D STORM imaging of Alexa Fluor 647 stained microtubules in a VECTASHIELD Mounting Medium/TRIS-Glycerol mixture. (Image kindly supplied by Dr. Nicolas Olivier, King's College, London UK.) VECTASHIELD® Antifade Mounting Medium Catalog #: H-1000
Single copy DNA probe to the Downs region of chromosome 21 using FITC-labeled antibody (green) and mounted in VECTASHIELD Mounting Medium with DAPI (blue) - Photo courtesy of Cytocell Ltd. VECTASHIELD® Antifade Mounting Medium with DAPI Catalog #: H-1200
FastTag Fluorescein labeled pUC1.77 detected with Biotinylated Anti-Fluorescein and Fluorescein Avidin DCS, and counterstained with VECTASHIELD with PI. VECTASHIELD® Antifade Mounting Medium with Propidium Iodide (PI) Catalog #: H-1300
Alzheimer's disease mouse brain model - double label: glial cells stained with Iba1 (rp) followed by DyLight 594 Goat Anti-Rabbit IgG (DI-1594); plaques stained with Biotinylated Mouse Anti-Human Amyloid Beta followed by DyLight 488 Streptavidin (SA-5488 VECTASHIELD® HardSet™ Antifade Mounting Medium Catalog #: H-1400
Mouse embryonic fibroblast stained with mouse anti-tubulin followed by fluorescein horse anti-mouse IgG and mounted in VECTASHIELD HardSet Mounting Medium with DAPI and TRITC-Phalloidin. VECTASHIELD® HardSet™ Antifade Mounting Medium with Phalloidin Catalog #: H-1600

Technical Information

VECTASHIELD Mounting Media are compatible with a wide array of fluorochromes, enzymatic substrates, and fluorescent proteins. Please consult the compatibility table (in the "Documents" section) to determine if VECTASHIELD will be compatible in your system.

All formulations of VECTASHIELD Antifade Mounting Media are available with or without DAPI (4', 6-diamidino-2-phenylindole) counterstain.  The DAPI concentration can be modified by mixing with the corresponding VECTASHIELD Mounting Medium without DAPI.  DAPI produces a blue fluorescence when bound to DNA with excitation at about 360 nm and emission at 460 nm.

R.J. Florijn, et. al. Cytometry, 19 (1995) 177-182.

The refractive index for VECTASHIELD HardSet Mounting Medium is 1.46.

NOTE: This product has a six month expiration date.

VECTASHIELD Mounting Medium Antifade Comparison

Other manufacturers measure the antifade properties of their mountants using labeled microspheres or arrayed spots. Vector Labs prefers to measure antifade properties of VECTASHIELD mountants using frozen tissue sections immunohistochemically stained with fluorescently labeled secondary antibodies. Antifade capability is measured using a 40x objective with real time imaging over 30 seconds of continuous exposure to the excitation illumination. Individual intensity measurements are recorded from 6 separate labeled regions and the average is calculated. The intensity after 30 second exposure is expressed as a percentage of the intensity at zero time. The values for PG are taken from the manufacturer s published results.

VECTASHIELD Anifade Comparison
VECTASHIELD HardSet Antifade Properties
 
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