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ExoBrite™ EV Total RNA Isolation Kit

Optimized and easy-to-use kit for extracting total RNA, including mRNA and miRNA, from purified EVs for downstream analysis in qPCR or RNAseq.

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Product Description

Obtaining high yields of quality RNA from extracellular vesicles (EVs) is a challenge due the dilute nature of EV samples and low RNA content within EVs. In addition, short non-coding RNAs within EVs, such as miRNAs, are commonly studied as biomarkers and often require specialized methods for efficient extraction. The ExoBrite™ EV Total RNA Isolation Kit was designed to address these challenges by offering an optimized and easy-to-use kit for total RNA isolation, including mRNA and miRNA, from purified EVs. The isolated EV RNA can then be used for downstream analysis such as qPCR or RNAseq.

Features

  • Extract total RNA from purified EVs
  • Recover ~10-20 ng of RNA from 1×1010 SEC-enriched EVs
  • Compatible with downstream applications such as qPCR or RNAseq
  • Simple column-based purification
  • No phenol/chloroform or ethanol precipitation steps

Kit components

  • EV Lysis Buffer
  • Column Binding Buffer
  • DNase Buffer
  • DNase I (lyophilized)
  • Column Wash Buffer
  • Spin Columns

The RNA isolation procedure is a simple column purification method that takes as little as 20 minutes and requires no phenol/chloroform or ethanol precipitation steps. The kit is estimated to recover
~10-20 ng of RNA from 1×1010 SEC-enriched EVs. The amount of RNA recovered will depend primarily on EV number and quality, which may be affected by purification method, storage conditions, and number of freeze thaws. An optional, but recommended, DNase treatment step is used to remove contaminating DNA.

See below our full catalog of ExoBrite™ stains and antibodies developed for EV analysis by flow cytometry, western blotting, or super-resolution imaging.

Full List of ExoBrite™ Products for EV Research

ProductConjugateDetection channelsSizeCatalog Number
ExoBrite™ True EV Membrane StainsN/AFITC, PE100 Labelings, 500 Labelings30129, 30130
ExoBrite™ Annexin EV Staining KitsAnnexin VPacific Blue™, FITC, PE, Cy®3, APC100 Labelings, 500 Labelings30119-30122
ExoBrite™ WGA EV Staining KitsWheat Germ Agglutinin (WGA)30123-30126
ExoBrite™ CTB EV Staining KitsCholera Toxin Subunit B (CTB)30111-30114
ExoBrite™ EV Surface Stain Sampler Kit, GreenAnnexin V, Wheat Germ Agglutinin (WGA), Cholera Toxin Subunit B (CTB)FITC100 Labelings30127
ExoBrite™ STORM CTB EV Staining KitsCholera Toxin Subunit B (CTB)FITC, Texas Red®, Cy®5, Cy®5.5100 Labelings, 500 Labelings30115-30118
ExoBrite™ CD9 Flow AntibodyExoBrite™ 410/450
ExoBrite™ 490/515
ExoBrite™ 560/585
ExoBrite™ R-PE
Pacific Blue ™
FITC
PE
25 tests
100 tests
P003-410... P003-RPE
ExoBrite™ CD63 Flow AntibodyP004-410... P004-RPE
ExoBrite™ CD81 Flow AntibodyP005-410... P005-RPE
ExoBrite™ IgG1 Isotype Control Flow AntibodyP008-410... P008-RPE
ExoBrite™ CD9 Western AntibodyExoBrite™ 680/700
ExoBrite™ 770/800
700 channel
Far-red channel
25 tests
100 tests
P003-680, P003-770
ExoBrite™ CD63 Western AntibodyP004-680, P004-770
ExoBrite™ CD81 Western AntibodyP006-680, P006-770
ExoBrite™ Calnexin Western AntibodyExoBrite™ 770/800800 channel
NIR channel
P007-770
ExoBrite™ Streptavidin Magnetic BeadsN/AN/A5 mL28000

Product Attributes

Size
50 preps

Documents, Protocols, SDS and COA

FAQs

Product shipping, storage, shelf life, & solubility

Bioscience kits
The guaranteed shelf life from date of receipt for bioscience kits is listed on the product information sheet. Some kits have an expiration date printed on the kit box label, this is the guaranteed shelf life date calculated from the day that the product shipped from our facility. Kits often are functional for significantly longer than the guaranteed shelf life. If you have an older kit in storage that you wish to use, we recommend performing a small scale positive control experiment to confirm that the kit still works for your application before processing a large number of samples or precious samples.

Antibodies and other conjugates
The guaranteed shelf life from date of receipt for antibodies and conjugates is listed on the product information sheet. Antibodies and other conjugates often are functional for significantly longer than the guaranteed shelf life. If you have an older conjugate in storage that you wish to use, we recommend performing a small scale positive control experiment to confirm that the product still works for your application before processing a large number of samples or precious samples.

For lyophilized antibodies, we recommend reconstituting the antibody with glycerol and antimicrobial preservative like sodium azide for the longest shelf life (note that sodium azide is not compatible with HRP-conjugates).

Chemicals, dyes, and gel stains
Biotium guarantees the stability of chemicals, dyes, and gel stains for at least a year from the date you receive the product. However, the majority of these products are highly stable for many years, as long as they are stored as recommended. Storage conditions can be found on the product information sheet or product safety and data sheet, material safety data sheet, and on the product label. Fluorescent compounds should be protected from light for long term storage.

If you have a Biotium compound that has been in storage for longer than one year that you wish to use, we recommend performing a small scale positive control experiment to confirm that the compound still works for your application before processing a large number of samples or precious samples.

Expiration date based on date of manufacture (DOM)
If your institution requires you to document expiration date based on date of manufacture for reagents, please contact techsupport@biotium.com for assistance.

Chemical products with special stability considerations:

Esters

Ester compounds include the following:

  • Succinimidyl esters (SE, also known as NHS esters), such as our amine-reactive dyes
  • Acetoxymethyl esters (AM esters), such as our membrane-permeable ion indicator dyes
  • Diacetate-modified dyes, like ViaFluor™ 405, CFDA, and CFDA-SE cell viability/cell proliferation dyes

Ester dyes are stable in solid form as long as they are protected from light and moisture. Esters are not stable in aqueous solution. Concentrated stock solutions should be prepared in anhydrous DMSO (see Biotium catalog no. 90082). Stock solutions in anhydrous DMSO can be stored desiccated at -20°C for one month or longer. Esters should be diluted in aqueous solution immediately before use. Succinimidyl esters (SE) should be dissolved in a solution that is free of amine-containing compounds like Tris, glycine, or protein, which will react with the SE functional group. AM esters and diacetate compounds should be dissolved in a solution that is free of serum, because serum could contain esterases that would hydrolyze the compound.

A note on CF® Dye succinimidyl ester stability

Succinimidyl esters (SE) are generally susceptible to hydrolysis, which can result in lower labeling efficiency. Many commercially available fluorescent dyes used for life science research are heavily sulfonated dyes which makes them particularly hygroscopic, worsening the hydrolysis problem. In addition, for several commercially available SE reactive dyes, the SE group is derived from an aromatic carboxylic acid, while the SE group in all of Biotium’s CF® Dyes is prepared from an aliphatic carboxylic acid. This structural difference reduces the susceptibility of CF® Dye SE reactive groups to hydrolysis, resulting in relatively stable reactive dyes with consistently higher labeling efficiency compared to other SE derivatives of other fluorescent dyes.

Maleimides, MTS and thiosulfate dyes
Like the succinimidyl ester dyes, these dyes are also susceptible to hydrolysis, although generally to a much lower degree. Thus, for long term storage, anhydrous DMSO is recommended for making stock solutions.

Other reactive dyes
Amines, aminooxy (also known as oxylamine), hydrazide, azide, alkyne, BCN, and tyramide reactive dyes, as well as dye free acids, are generally stable in aqueous solution when stored at -20°C for 6-12 months or longer, as long as no compounds are present that may react with the dye’s functional group. See the product information sheets for specific reactive dyes more information.

Coelenterazines and D-luciferin

Coelenterazines are stable in solid form when stored as recommended; they are not stable in aqueous solution. Concentrated coelenterazine stock solutions (typically 1-100 mg/mL) should be prepared in ethanol or methanol; do not use DMSO or DMF to dissolve coelenterazines, because these solvents will oxidize the compounds. Ethanol or methanol stocks of coelenterazine can be stored at -20°C or below for six months or longer; alcohol stocks may evaporate during storage, so use tightly sealing screw cap vials and wrap the vials with Parafilm for long term storage. Propylene glycol also can be used as a solvent to minimize evaporation. If the solvent evaporates, the coelenterazine will still be present in the vial, so note the volume in the vial prior to storage so that you can adjust the solvent volume to correct for evaporation if needed. Prepare working solutions in aqueous buffers immediately before use. Coelenterazines are stable for up to five hours in aqueous solution.

Aquaphile™ coelenterazines are water soluble formulations of coelenterazines. They are stable in solid form when stored as recommended. Aquaphile™ coelenterazines should be dissolved in aqueous solution immediately before use. They are stable for up to five hours in aqueous solution.

Note that coelenterazines are predominantly yellow solids, but may contain dark red or brown flecks. This does not affect product stability or performance. If your coelenterazine is uniformly brown, then it is oxidized and needs to be replaced.

D-luciferin is stable in solid form and as a concentrated stock solution when stored as recommended; it is not stable at dilute working concentrations in aqueous solution. Prepare concentrated D-luciferin stock solutions (typically 1-100 mg/mL) in water, and store in aliquots at -20°C or below for six months or longer. Prepare working solutions immediately before use.

For dyes or reagents that are supplied lyophilized (as solids), it is hard to compare quantities based on appearance of the dye in the tube, because during the lyophilization process the dye can dry down in different ways, either spread out all over the tube, clumped together, or coating the sides or bottom of the tube. Centrifugation of the tube may not help in collecting the dye solid to the bottom of the tube as this generally works for solutions. However, lyophilized solids are packaged based on highly accurate absorbance measurement of the reagent solution prior to drying, so the vial will contain the correct amount of dye.

Biotium ships all antibodies (primary, secondary and conjugates) at room temperature. We guarantee their quality and performance under these conditions based upon our stability testing. Antibodies were subjected to accelerated stability testing by storing them at various temperatures (4°C, room temperature, or 37°C) for 1 week to mimic simulated shipping conditions and tested in immunostaining experiments. All antibodies showed the expected brightness and specificity, even after storage at sub-optimal temperatures for a week or longer. You can also download our Product Storage Statement here.

In line with our goal to be more environmentally friendly by reducing the use of excess packaging, and lowering shipping costs for our customers, products that have passed our stability testing are shipped at room temperature.

Once you have received the antibody vial, please follow the long-term storage instructions on the product information (PI) sheet.

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