TIGIT is an immune receptor present on some T cells and Natural Killer cells. TIGIT binds with high affinity to the poliovirus receptor (PVR) which causes increased secretion of IL10 and decreased secretion of IL12B and suppresses T cell activation by promoting the generation of mature immunoregulatory dendritic cells. Through the CD226/TIGIT-PVR pathway, TIGIT regulates T cell mediated immunity. In cancer, TIGIT and PD-1 have been shown to be over-expressed on tumor antigen-specific CD8+ T cells and CD8+ tumor infiltrating lymphocytes (TILs) from individuals with melanoma. Blockade of TIGIT and PD-1 led to increased cell proliferation, cytokine production, and degranulation of tumour antigen-specific CD8+ T cells and TIL CD8+ T cells. It can be considered an immune checkpoint.
This product is suitable for rapid extraction of DNA from tissue, cells, blood, saliva, swabs, blood spots, semen and other clinical samples. DNA can be used directly for PCR, quantitative PCR, Southern Blot, test of virus DNA and so on.
Specifications
| Features | Specifications |
| Main Functions | Isolation total DNA from blood, buffy coat, tissue and other samples |
| Applications | Second generation sequencing, PCR, real time PCR, etc. |
| Purification method | Polydisperse magnetic beads |
| Purification technology | Magnetic beads technology |
| Process method | Manual or automatic |
| Sample type | Anticoagulant blood, concentrated blood, buffy coat, lymphocytes and cultured cells |
| Sample amount | Whole blood :< 200μl; Saliva / swab:< 400μl; Tissue :< 20mg |
| Yield | 0.1 – 50μg |
| Elution volume | |
| Time per run |
This product is based on the purification method of high binding magnetic particles. The sample is lysed and digested under the action of lysate and protease. DNA is released into the lysate. After adding magnetic particles and binding solution, DNA will be adsorbed on the surface of magnetic particles, and impurities such as proteins will be removed without adsorption. The adsorbed particles were washed with washing solution to remove proteins and impurities, washed with ethanol to remove salts, and finally DNA was eluted by Elution Buffer.
| Contents | IVD3102 |
| Purification Times | 200 |
| MagPure Particles | 5 ml |
| Proteinase K | 100 mg |
| Protease Dissolve Buffer | 10 ml |
| Rnase A | 40 mg |
| Buffer ATL | 60 ml |
| Buffer AL | 60 ml |
| Buffer BD* | 20 ml |
| Buffer BW1* | 110 ml |
| Elution Buffer | 30 ml |
| Cat.No | Reagent | IVD3102-F-96 |
| Proteinase K | 50 mg | |
| Protease Dissolve Buffer | 6 ml | |
| RNase A | 20 mg | |
| Buffer ATL | 30 ml | |
| Buffer AL | 30 ml | |
| 96-Tip | 1 | |
| Sample plate (DW Plate) | 450µl Buffer BD(Ethanol Added) | 1 |
| Wash 1 Plate (DW Plate) | 600µl Buffer BW1(Ethanol Added) | 1 |
| Wash 2 Plate (DW Plate) | 600µl Buffer BW1(Ethanol Added) | 1 |
| Wash 3 Plate (DW Plate) | 750µl Buffer GW2, 20µl MagPure Particle | 1 |
| Elution plate (DW Plate) | 80µl Elution Buffer | 1 |
| Cat.No | Reagent | IVD3102-TL-06 |
| Proteinase K | 50 mg | |
| RNase A | 20 mg | |
| Protease Dissolve Buffer | 6 ml | |
| Buffer ATL | 40 ml | |
| Buffer AL | 40 ml | |
| AS-Tip | 12 | |
| 2.0ml V-bottom plate | Row 1/7：450µl Buffer BDRow 2/8：450µl Buffer BW1Row 3/9：450µl Buffer BW1Row 4/10：20μl Magpure Particle450μl Wash Buffer GW2 Row 5/11：450μl Wash Buffer GW2 Row 6/12：80µl Elution Buffer | 6 |
Storage and Stability
Proteinase K, RNase A, MagPure Particles should be stored at 2-8°C upon arrival. However, short-term storage (up to 24 weeks) at room temperature (15-25°C) does not affect their performance. The remaining kit components can be stored at room temperature (15-25°C) and are stable for at least 18 months under these conditions.
Experiment Data
This product is suitable for rapid extraction of DNA from tissue, cells, blood, saliva, swabs, blood spots, semen and other clinical samples. DNA can be used directly for PCR, quantitative PCR, Southern Blot, test of virus DNA and so on.
The amount of RNA that can be extracted from different biological or clinical samples varies greatly. For example, while a few micrograms of RNA could be easily purified from tissues and cells in excess amounts (such as from a few milligrams of tissue), many liquid biopsy samples may yield very low amounts of RNA. In fact, samples such as urine or plasma may yield 1 – 100 ng or less RNA per 100 µL of sample. Such a range of RNA quantity is often below the detection limit of most commonly used techniques for measuring RNA including nano-spectrophotometry and fluorescent nucleic acid stains. As a result, without properly determined RNA concentration, it becomes very difficult to normalize the starting quantity of RNA used in gene expression studies.
Norgen’s microRNA (cel-miR-39) Spike-In Kit offers a quantified synthetic RNA (cel-miR-39) for spike-in during RNA extraction procedures and subsequent normalization in RT-qPCR assays. The amount of cel-miR-39 RNA recovered after RNA extraction is directly correlated with the amount of total RNA recovered. After reverse transcription (such as with Norgen’s microScript Reverse Transcription system) of the sample RNA (with spike-in), the level of cel-miR-39 can be determined by subjecting the cDNA generated to quantitative PCR (qPCR) using fluorescent nucleic acid stains such as SYBR Green. A cel-miR-39 specific primer is included in the kit. The level of expression of any target transcripts in different RNA samples can now be normalized to the cel-miR-39 transcript level using standard method such as ∆∆Ct relative quantification.
In addition, the cel-miR-39 RNA is compatible to library preparation methods (including ligation-based protocols) in Next Generation Sequencing (Small RNA-Seq) workflows. The cel-miR-39 RNA could be used for normalization as well as for tracking library construction efficiency.
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Storage Conditions
Upon receipt, store Norgen’s microRNA (cel-miR-39) Spike-In Kit at -20°C or lower. Avoid multiple freeze-thaw cycles. If needed, prepare smaller working aliquots and store at -20°C or lower.
| Component | Cat. 59000 |
|---|---|
| cel-miR-39 RNA | 10 pmol |
| cel-miR-39 Forward PCR Primer | 1 nmol |
| Nuclease-Free Water | 1.25 mL |
| Product Insert | 1 |
Norgen’s Plasma/Serum Exosome and RNA Isolation Kits constitute all-in-one systems for the purification of exosomes and the sequential isolation of exosomal 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, irrespective of size or GC content, without bias. 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. The RNA isolated from the purified exosomes is free from any protein-bound circulating RNA and is of the highest integrity. 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.
Norgen’s Plasma/Serum Exosome and RNA Isolation Mini Kit
For sample volumes ranging from 50 µL to 1 mL. This kit allows the user to elute into a flexible elution volume ranging from 50 µL to 100 µL.
Norgen’s Plasma/Serum Exosome and RNA Isolation Midi Kit
For sample volumes ranging from 1 mL to 4 mL. This kit allows the user to elute into a flexible elution volume ranging from 50 µL to 100 µL.
Norgen’s Plasma/Serum Exosome and RNA Isolation Maxi Kit
For sample volumes ranging from 4 mL to 10 mL. This kit allows the user to elute into a flexible elution volume ranging from 50 µL to 100 µL.
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| Kit Specifications | |
| Minimum Plasma/Serum Input | 4 mL |
| Maximum Plasma/Serum Input | 10 mL |
| Size of Exosomes Purified | 40 nm – 150 nm |
| Size of RNA Purified | All sizes, including miRNA and small RNA (< 200 nt) |
| Elution Volume | 50-100 μL |
| Time to Complete 10 Purifications | 35 – 40 minutes |
| Average Yields* | Variable depending on specimen |
*Please check page 4 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. 58300 (50 preps) | Cat. 58500 (25 preps) | Cat. 58600 (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 | 20 mL | 20 mL | 20 mL |
| Lysis Additive B | 2 mL | 2 mL | 2 mL |
| Wash Solution A | 38 mL | 18 mL | 18 mL |
| Elution Solution A | 6 mL | 6 mL | 6 mL |
| Mini Filter Spin Columns | 50 | 25 | 15 |
| Mini Spin Columns | 50 | 25 | 15 |
| Collection Tubes | 50 | 25 | 15 |
| Elution Tubes (1.7 mL) | 100 | 50 | 30 |
| Product Insert | 1 | 1 | 1 |
