APExBIO Technology LLC 品牌 ->>肝素管柱 (HyperTrap Heparin HP Column)

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雷帕黴菌（Rapamycin）| Romidepsin (FK228, depsipeptide) | Daptomycin | G-418 (80 S 核醣體延伸抑制劑) 

肝素管柱 (HyperTrap Heparin HP Column) APExBIO  貨號PC1009

Heparin affinity chromatography preloaded column with high performance, resistance to organic solvents and chemical stability.

HyperChrom HP肝素瓊脂糖 (HyperChrom Heparin HP Agarose) 是一種親和性層析介質(chromatography medium )，由肝素共價偶聯(covalently coupling heparin)到瓊脂糖基質(agarose base)上而成。肝素( Heparin)是一種天然存在的糖胺聚醣( glycosaminoglycan)，與多種生物分子具有親和力，也可用作離子交換配體( ion exchange ligand)。此層析介質可用於分離和純化凝血因子(coagulation factors)、抗凝血酶 III(antithrombin III)、生長因子( growth factors)、干擾素(interferon)、脂蛋白脂肪酶(lipoprotein lipase)以及針對核酸(nucleic acid )和類固醇受體( steroid receptors)的酵素。與 HyperChrom FF肝素瓊脂糖(HyperChrom Heparin FF Agarose)相比，HyperChrom HP肝素瓊脂糖 (HyperChrom Heparin HP Agarose) 粒徑更細，解析度更高。HyperTrap 肝素 HP 層析管採用 HyperChrom 肝素 HP 瓊脂糖作為層析介質。預裝柱管主體及內塞採用PP材質，內外拋光，篩板採用HDPE材質，具備優良的物理性能及耐化學腐蝕性，耐腐蝕、抗老化、壽命長、安全性佳。 即用型( Ready-to-use)，方便快速。 可與注射器( syringes)、蠕動幫浦或層析系統搭配使用。 可串聯使用，增加樣品處理能力。

HyperTrap Heparin HP Column preloaded column parameters

HyperTrap Heparin HP Column chromatography media parameters

2. Sample preparation
Prepare samples for purification.

3. Chromatographic conditions
Flow rate selection: Linear flow rate of 90-150 cm/h is generally selected according to the height of the column bed.
Sample preparation: To prevent the sample from clogging the column, the sample needs to be filteredwith a 0.2/0.45 μm (after inclusion body disruption) microporous membrane before loading, and it is recommended that the pH and conductivity of the sample be adjusted to be consistent with the equilibrium buffer

Column efficiency determination (optional).

Select one of the two test methods shown in the table below for column effectiveness testing. Use the
mobile phase equilibrium chromatography column to the baseline to be stable, load the sample into the
chromatography column, continue to use the mobile phase for rinsing, and after the chromatographic peak
is completed to return to the baseline, end the run, integrate the chromatographic peak, and evaluate the
loading effect

Preloaded column setup
(1) Open the package and remove the preloaded column.
(2) Connect the pre-packed column: unscrew the upper and lower plugs of the pre-packed column and connect it to the chromatography system (peristaltic pump or syringe, etc.), rinse the pre-packed column with pure water 2-3 times the column volume to drain the preservation solution (generally 20% ethanol). In order to prevent air bubbles from entering, pure water can be dripped and filled at the inlet end of the pre-loaded column before connecting the system connector.
(3) Cleaning and disinfection: For the first use, it is recommended to clean and disinfect the column and rinse 2 times the column volume with pure water or buffer. For recommended cleaning and
disinfection buffers, refer to subsequent cleaning and regeneration procedures.
(4) Sample preparation: To prevent the sample from clogging the column, it is recommended that the sample be filtered with a 0.45 μm microporous membrane before loading

6. Chromatographic steps

(1) Equilibrium: Use Balance/Bind/Wash Buffer to fully equilibrate the column to pH and conductivity stable and substantially consistent with the equilibration buffer, which typically requires 3-5 times the column volume.
(2) Sample loading: Determine the sample loading volume and amount on the Hyper Chrom Heparin HP Agarose based on the binding load measured in the pilot experiment.
(3) Washing: Rinse the column with Balance/Bind/Wash Buffer or other suitable buffers until UV stable and return to baseline.
(4) Elution: Elution is achieved by increasing the concentration of salt ions, which can be gradually increased by linear gradient or step gradient to elute molecules with different binding strengths.
(5) Regeneration: Rinse the column with buffer containing high salt such as 2 M NaCl.
(6) Rebalancing: Re-equilibrate the chromatography column with Balance/Bind/Wash Buffer.

7. Cleaning and recycling
As the number of uses of the chromatography medium increases, contaminants (e.g., lipids, endotoxins, proteins, etc.) accumulate on the chromatography column. Regular in-place cleaning is essential to keep the column in stable working condition. Determine the frequency of in-place cleaning according to the degree of contamination of the chromatography medium (if the contamination is serious, it is recommended that in-place cleaning should be carried out after each use to ensure repeatable results and extend the working life of the chromatography medium).
For different types of impurities and contaminants, cleaning can be carried out under the following conditions:

Removal of strongly binding proteins: Wash with 2 M NaCl solution in 5x column volume.
Denatured and removal of precipitated proteins: first wash with 0.1 M NaOH solution in 5x column volume, then wash the lye with 5-10 column volumes of purified water. It can also be
washed with 6 M guanidine hydrochloride or 8 M urea.

Removal of hydrophobic and lipid substances: 0.1-0.5% nonionic detergent washed followed by
rinse with 5-10 column volumes of purified water.
Note: The flow rate can be selected from 30-60 cm/h during the cleaning process; When the
blockage is serious, reverse cleaning can be used.

8. sterilization
In order to reduce the microbial load, it is recommended to use 0.5~1 M NaOH solution to treat the
chromatography medium with a processing time of 15~30 min.

9. stockpile

(1) Equilibrium: Use Balance/Bind/Wash Buffer to fully equilibrate the column to pH and conductivity stable and substantially consistent with the equilibration buffer, which typically requires 3-5 times the column volume.
(2) Sample loading: Determine the sample loading volume and amount on the Hyper Chrom Heparin HP Agarose based on the binding load measured in the pilot experiment.
(3) Washing: Rinse the column with Balance/Bind/Wash Buffer or other suitablebuffers until UV stable and return to baseline.
(4) Elution: Elution is achieved by increasing the concentration of salt ions, which can be gradually increased by linear gradient or step gradient to elute molecules with different binding strengths.
(5) Regeneration: Rinse the column with buffer containing high salt such as 2 M NaCl.
(6) Rebalancing: Re-equilibrate the chromatography column with Balance/Bind/Wash Buffe

Unopened chromatography media, please keep in the original container; The completed chromatography column is first soaked with 20% ethanol solution and then the upper and lower column heads are closed. The storage environment is 4~30oC.

10. Destruction and recycling
Since Hyper Chrom Heparin HP Agarose chromatography media is difficult to degrade in nature, incineration of discarded chromatography media is recommended in order to protect the environment. For chromatography media exposed to bioactive samples such as viruses and blood, please follow local biosafety requirements before destroying or disposing of them

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APExBIO提供優質的胜肽和生物分析試劑，廣泛產品線如Protease、Chromatin/Epigenetics表觀遺傳學、MAPK Signal、DNA Damege/DNA repair、細胞凋亡、腫瘤生物學、Stem cell等研究領域， 還提供定制的服務，包括多肽合成，抗體的生產和檢測的發展。產品線DNA/RNA prep kit、Drug Screening panel、Bioactive peptides、Growth factors、Biotinylation Reagents、tag peptides、amino Acids、Growth factors等。APExBIO產品質量都伴隨著證書的分析，高效液相色譜，質譜，和HMNR數據。

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