Norgen’s FBS Exosome Depletion Kits provides a quick and easy protocol for the depletion of bovine exosomes from FBS prior to using it as a growth supplement in your culture medium. The FBS recovered from the depletion process is exosome-depleted and does not contain any quantifiable bovine miRNAs. Moreover, the exosome-depleted FBS will support the growth of your cells of interest similar to the non-depleted FBS. Norgen’s kits allow for the processing of different FBS volumes. The depletion is based on Norgen’s proprietary resin. These kits provide a clear advantage over other available kits in that they do not require ultracentrifugation, any special instrumentation, precipitation reagents or any protease treatments. More importantly, the depletion process is an inexpensive method for exosome depletion from FBS, as compared to the expensive current ready-to-use exosome-depleted media available on the market.
Background
Most culture medium used for the growth and propagation of cells in culture require the addition of fetal bovine serum (FBS) as a growth supplement to media. FBS is obtained from bovine (cow) serum, and therefore contains large quantities of bovine exosome vesicles. These exosomes may interfere with some types of studies, or may lead to unreliable results when studying the exosomes shed from your cells of interest in normal culture conditions. Therefore, the use of exosome-depleted FBS is highly recommended for many types of studies.
FBS Exosome Depletion Kit (Slurry Format)
For FBS volumes ranging from 140 mL to 280 mL.
FBS Exosome Depletion Kit (Column Format)
For FBS volumes ranging from 120 mL to 240 mL.
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Kit Specifications | |
Sample Type | Fetal Bovine Serum |
Sample Volume Range | Up to 140 mL (FBS Exosome Depletion Kit I (Slurry Format) Up to 280 mL (FBS Exosome Depletion Kit II (Slurry Format) |
Depletion | Deplete exosome-sized vesicle |
Bovine miRNA | No detectable bovine miRNA |
Time to Complete 6 Purifications | 40 minutes |
Storage Conditions and Product Stability
All buffers should be kept tightly sealed and stored at room temperature. This kit is stable for 2 years after the date of shipment.
Component | Cat. 61100 (6 preps) | Cat. 61400 (12 preps) | Cat. 61200 (6 preps) | Cat. 61300 (12 preps) |
---|---|---|---|---|
ExoC Buffer | 2 x 1.5 mL | 8 mL | 2 x 1.5 mL | 8 mL |
Slurry E | 12.5 mL | 2 x 12.5 mL | – | – |
Maxi Spin Column | – | – | 6 | 12 |
Product Insert | 1 | 1 | 1 | 1 |
These kits provide a fast, reliable and convenient method to sequentially isolate and concentrate exosomal RNA as well as Free-Circulating RNA from different plasma/serum sample volumes. The purification is based on spin column chromatography that employs Norgen’s proprietary resin. These kits are designed to isolate all sizes of RNA, including microRNA as well as all sizes of the free-circulating protein-bound RNA, including microRNA. These kits provide a clear advantage over other available kits in that they do not require any special instrumentation, protein precipitation reagents, extension tubes, phenol/chloroform or protease treatments. Moreover, these kits allow the user to elute into a flexible elution volume ranging from 50 µL to 100 µL. The RNA isolated from the purified exosomes is free from any protein-bound circulating RNA and is of the highest integrity. Moreover, the free-circulating, protein-bound, RNA is free from any exosomal RNA. The purified RNA can be used in a number of downstream applications including real time PCR, reverse transcription PCR, Northern blotting, RNase protection and primer extension, and expression array assays.
Plasma/Serum Exosome and Free-Circulating RNA Isolation Mini Kit
For sample volumes ranging from 50 µL to 1 mL.
Plasma/Serum Exosome and Free-Circulating RNA Isolation Midi Kit
For sample volumes ranging from 1 mL to 4 mL.
Plasma/Serum Exosome and Free-Circulating RNA Isolation Maxi Kit
For sample volumes ranging from 4 mL to 10 mL.
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Kit Specifications | |
Minimum Plasma Input | 1 mL |
Maximum Plasma Input | 4 mL |
Size of RNA Purified | All sizes, including miRNA and small RNA (< 200 nt) |
Elution Volume | 50 – 100 µL |
Time to Complete 10 Purifications | 40 – 45 minutes |
Average Yields* | Variable depending on specimen |
*Please check page 5 of the product insert for the average yields and the common RNA quantification methods
Storage Conditions and Product Stability
All solutions should be kept tightly sealed and stored at room temperature. This kit is stable for 2 years after the date of shipment. It is recommended to warm Lysis Buffer A for 20 minutes at 60°C if any salt precipitation is observed.
Important Note
This kit is suitable for the purification of exosomes from fresh or frozen serum or plasma prepared from blood collected on either EDTA or Citrate. Plasma samples prepared from blood collected on heparin should not be used as heparin can significantly interfere with many downstream applications such as RT-PCR.
Component | Cat. 59500 (50 preps) | Cat. 59600 (25 preps) | Cat. 59700 (15 preps) |
---|---|---|---|
Slurry E | 12.5 mL | 12.5 mL | 12.5 mL |
ExoC Buffer | 8 mL | 8 mL | 2 x 8 mL |
ExoR Buffer | 12 mL | 12 mL | 12 mL |
Lysis Buffer A | 2 x 20 mL | 2 x 20 mL | 2 x 20 mL |
Lysis Additive B | 2 mL | 2 mL | 2 mL |
Wash Solution A | 2 x 18 mL | 18 mL | 18 mL |
Elution Solution A | 2 x 6 mL | 6 mL | 6 mL |
Mini Filter Spin Columns | 50 | 25 | 15 |
Mini Spin Columns | 100 | 50 | 30 |
Collection Tubes | 100 | 50 | 30 |
Elution Tubes (1.7 mL) | 100 | 50 | 30 |
Product Insert | 1 | 1 | 1 |
Clone | IHC623 |
Source | Mouse Monoclonal |
Positive Control | Breast, Colon, Associated Adenocarcinomas |
Dilution Range | 1:200 |
Mucin 1 (MUC1) is a membrane-bound glycoprotein involved in a number of protective and cell-signaling functions, including cell-cell adhesion, proliferation, motility, invasion, and survival. Overexpression of MUC1 is clinically indicated in breast carcinomas, papillary thyroid carcinomas, and thymic carcinomas, and reports have named MUC1 as a useful marker for differentiating thymic carcinoma from type B3 thymoma. The expression of MUC1 is correlated with the grade of malignancy in thymic epithelial tumors, and loss of MUC1 expression has been associated with reactive gastropathy. MUC1 is not expressed in normal human epidermis, but has been detected in the epidermis of psoriatic plaques of biopsies from patients diagnosed with psoriasis vulgaris.
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