Nerve Growth Factor Receptor (NGFR), also known as p75, P-75NTR or CD271, is a neurotrophin receptor belonging to the tumor necrosis factor receptor family. NGFR is expressed mainly in Schwann cells and neurons, as well as a number of other non-neuronal cell types, and functions during central and peripheral nervous system development to regulate neuronal growth, migration, differentiation, and cell death. Nerve Growth Factor Receptor is also expressed in melanocytes, melanomas, neuroblastomas, pheochromocytomas, neurofibromas, neurotized nevi (type C melanocytes), and other neural crest cell or tumor derivatives. It has been suggested that NGFR may act as a tumor suppressor indicated in prostate and urothelial cancer, and Anti-Nerve Growth Factor Receptor (NGFR) is often used in adjunct with S100, to aid in the diagnosis of desmoplastic and neurotrophic malignant melanomas. Anti-NGFR is also useful as an aid in the diagnosis of breast malignancy, as the antibody labels the myoepithelial cells of breast ducts and intralobular fibroblasts of breast ducts.
endo-BCN-PEG4-t-butyl ester is a click chemistry linker containing a BCN group and a t-butyl protected carboxyl group. The BCN group can react with azide-tagged molecules. The protected carboxyl group (COOH) prevents self coupling or polymerization under standard acid/amine or acid/hydroxyl coupling conditions. The t-butyl ester can be converted to free acid under acidc condition. Reagent grade, for research purpose. Please contact us for GMP-grade inquiries.
endo-BCN-PEG4-t-butyl ester is a click chemistry linker containing a BCN group and a t-butyl protected carboxyl group. The BCN group can react with azide-tagged molecules. The protected carboxyl group (COOH) prevents self coupling or polymerization under standard acid/amine or acid/hydroxyl coupling conditions. The t-butyl ester can be converted to free acid under acidc condition. Reagent grade, for research purpose. Please contact us for GMP-grade inquiries.
These kits provide a fast, reliable and convenient method to purify and concentrate high quality, high purity and inhibitor-free cell-free circulating and exosomal RNA using a convenient spin column method. These kits can purify RNA from fresh or frozen serum or plasma samples 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. The purified plasma/serum RNA is fully compatible with all downstream applications including PCR, qPCR, methylation-sensitive reverse transcription qPCR, reverse transcription PCR, Northern blotting, RNase protection and primer extension, expression array assays, and NGS. The EXTRAClean columns undergo stringent processing and rigorous quality control measures to minimize contamination traces, ensuring optimal results for sensitive applications such as NGS.
Cell-free circulating RNA, including exosomal RNA in plasma or serum, has the potential to provide biomarkers for certain cancers and disease states, and includes tumor-specific extracellular RNA in the blood. Exosomes are 40 – 100 nm membrane vesicles, which are secreted by most cell types. Exosomes can be found in saliva, blood, urine, amniotic fluid and malignant ascitic fluids, among other biological fluids. Evidence has been accumulating recently that these vesicles act as cellular messengers, conveying information to distant cells and tissues within the body. The exosomes contain cell-specific proteins, lipids and RNAs, which are transported to other cells, where they can alter function and/or physiology. These exosomes may play a functional role in mediating adaptive immune responses to infectious agents and tumours, tissue repair, neural communication and transfer of pathogenic proteins. Recent work has demonstrated the presence of distinct subsets of microRNAs within exosomes which depend upon the tumour cell type from which they are secreted. For this reason, exosomal RNAs may serve as biomarkers for various diseases including cancer. As the RNA molecules encapsulated within exosomes are protected from degradation by RNAses, they can be efficiently recovered from biological fluids, such as plasma or serum.
This kit can purify RNA from fresh or frozen serum or plasma samples prepared from blood collected on either EDTA or Citrate, from volumes ranging from 50 μL to 200 μL. The purified plasma/serum RNA is eluted in a flexible final volume of 10 μL to 25 μL.
This utilizes a two-column method, and can purify RNA from fresh or frozen serum or plasma samples prepared from blood collected on either EDTA or Citrate, from volumes ranging from 250 μL to 1.5 mL. The first column will handle the large volume input of bodily fluids that is followed by a concentration on a mini column for a final elution of 50 μL to 100 μL.
This kit can purify RNA from fresh or frozen serum or plasma samples prepared from blood collected on either EDTA or Citrate, from volumes ranging from 2 mL to 5 mL. The first column will handle the large volume input of bodily fluids that is followed by a concentration on a mini column for a final elution of 50 μL to 100 μL.
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| Kit Specifications | |
| Sample Type | Plasma/Serum |
| Anti-Coagulant (for Plasma)† | EDTA or Citrate |
| Sample Volume Range | 50 to 200 μL |
| Minimum Elution Volume | 10 μL |
| Maximum Elution Volume | 25 μL |
| Time to Complete 10 Purifications | 15 – 20 minutes |
| Size of RNA Purified | All sizes, including miRNA and small RNA (<200 nt) |
| Average Yields¥ | Variable depending on specimen |
† This kit is suitable for the isolation of RNA 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
¥ Please check page 5 for Average Plasma/Serum Yields and 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.
| Component | Cat. 73700 (50 preps) | Cat. 73710 (20 preps) | Cat. 73720 (10 preps) |
|---|---|---|---|
| Lysis Buffer A | 2 x 20 mL | 100 mL | 1 x 130 mL 1 x 30 mL |
| Wash Solution A | 18 mL* | 1 x 38 mL* 1 x 18 mL* | 38 mL* |
| Elution Solution A | 6 mL | 6 mL | 6 mL |
| Elution Buffer F | – | – | 15 mL |
| EXTRACLean Micro Spin Columns | 50 | – | – |
| EXTRACLean Mini Spin Columns | – | 20 | 10 |
| EXTRACLean Midi Spin Columns | – | 20 | – |
| EXTRACLean Maxi Spin Columns | – | – | 10 |
| Collection Tubes | – | 20 | 10 |
| Elution Tubes (1.7 mL) | 50 | 20 | 10 |
| Product Insert | 1 | 1 | 1 |
The HiPure Minipreps system provides a fast, simple, and cost-effective plasmid DNA miniprep method for routine molecular biology laboratory applications. HiPure Miniprep Kits use silica membrane technology to eliminate the cumbersome steps associated with loose resins or slurries. Plasmid DNA purified with Mini Kits is immediately ready for use. Phenol extraction and ethanol precipitation are not required, and high quality plasmid DNA is eluted in a small volume of Tris buffer or water.
Specifications
| Features | Specifications |
| Main Functions | Isolation up to 30μg endotoxin-free plasmid DNA from 1-5ml bacterial culture using 96-well bind plate |
| Applications | Cell transfection, animal injection, etc. |
| Purification method | 96 well plate |
| Purification technology | Silica technology |
| Process method | Manual (centrifugation or vacuum) |
| Sample type | Low copy plasmid vector |
| Sample amount | 1-5ml LB（x96） |
| Yield | 30μg |
| Elution volume | ≥75μl |
| Time per run | ≤60 minutes |
| Liquid carrying volume per column | 800μl |
| Binding yield of column | 70μg |
The HiPure Plasmid procedure is based on alkaline lysis of bacterial cells followed by adsorption of DNA onto silica in the presence of high salt. The unique silica membrane used in the kit completely replaces glass or silica slurries for plasmid DNA minipreps. The procedure consists of 3 basic steps: Preparation and clearing of a bacterial lysate by alkaline method，then transfer the supernatant to column to bind DNA. After washing proteins and other impurities, nucleic acid was finally eluted with low-salt buffer (10mm Tris, pH9.0, 0.5mm EDTA).
Advantages
1. High throughput: processing 96 samples at once;
2. High yield: The purified plasmid yield can reach up to 30μg, which is greater than the yield of the magnetic bead method kit;
3. Detoxification: endotoxin content <0.1EU/μg;
4. Fast: The entire extraction can be completed in 60 minutes without the need for time-consuming alcohol precipitation;
Kit Contents
| Contents | P115701 | P115702 | P115703 |
| Purification Times | 1 x 96 Preps | 4 x 96 Preps | 20 x 96 Preps |
| RNase A | 10 mg | 20 mg | 100 mg |
| Buffer P1 | 30 ml | 120 ml | 550 ml |
| Buffer P2 | 30 ml | 120 ml | 550 ml |
| Buffer LEN3 | 15 ml | 60 ml | 300 ml |
| Buffer LN4 | 80 ml | 270 ml | 2 x 700 ml |
| Buffer LN5 | 40 ml | 220 ml | 1100 ml |
| Buffer PW1 | 40 ml | 220 ml | 1100 ml |
| Buffer PW2 | 25 ml | 100 ml | 2 x 200 ml |
| Elution Buffer | 30 ml | 60 ml | 250 ml |
| Lysate Clear Plate | 1 | 4 | 20 |
| HiPure DNA Plate | 1 | 4 | 20 |
| 1.6ml Collection Plate | 2 | 8 | 40 |
| 0.5ml Collection Plate | 1 | 4 | 20 |
Storage and Stability
The kit components can be stored dry at room temperature (15-25°C) and are stable for at least 18 months under these conditions. If any precipitates form in the buffers, warm at 37℃ to dissolve. After addition of RNase A，Buffer P1 is stable for 6 months when stored at 2-8°C.
For any technical problems or customized products, please contact us.
F&Q about Endotoxin-free Plasmid Extraction Kit — P1156 ←click here
The HiPure Minipreps system provides a fast, simple, and cost-effective plasmid DNA miniprep method for routine molecular biology laboratory applications. HiPure Miniprep Kits use silica membrane technology to eliminate the cumbersome steps associated with loose resins or slurries. Plasmid DNA purified with Mini Kits is immediately ready for use. Phenol extraction and ethanol precipitation are not required, and high quality plasmid DNA is eluted in a small volume of Tris buffer or water.
