納米金粒子技術(shù)手冊（一）
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基于應用的黃金納米粒子產(chǎn)品選擇
| Application | Gold Nanoparticle Size Range |
Surface Chemistry | Benefits | Related Documents |
| Protein Conjugation | 5nm-100nm | standard (citrate) | Quick | Classic Passive Adsorption of Proteins to Gold Nanoparticles |
| ? | ? | NHS | Covalent conjugation to primary amines, increased stability, less non-specific protein binding. | Conjugation of proteins to NHS-activated gold nanoparticles |
| ? | ? | Maleimide | Covalent conjugation to thiol groups, increased stability, less non-specific protein binding. | - |
| ? | ? | Carboxyl | Covalent conjugation, increased stability, less non-specific protein binding. | Covalent conjugation of proteins to carboxyl gold nanoparticles |
| ? | ? | Amine | Conjugation of carboxylated ligands | - |
| ? | ? | Streptavidin | Can be used with any biotinylated ligand, ideal for high-throughput screenings. | - |
| Modification with thiolated ligands? (PEG-SH etc.) |
5nm-100nm | standard (citrate) | Classic starting material, no additional stabilizers added | - |
| ? | ? | stabilized (surfactant) | Increased stability during functionalization but reduced kinetics | - |
| Oligonucleotide Conjugation | 5nm-15nm | standard (citrate) | Ideal for conjugation of thiolated oligonucleotides to small particle sizes. | - |
| ? | 5nm-100nm | OligoREADY? | Ideal for conjugation of thiol modified oligos to particles between 5nm-100nm in diameter. | - |
| ? | 5nm-100nm | Maleimide | Ideal for covalent conjugation of thiol modified oligos to particles between 5nm-100nm in diameter. | - |
| ? | 5nm-100nm | NHS | For covalent conjugation of amine functionalized oligonucleotides. Ideal when a linker is required between the gold surface and conjugated oligonucleotide. |
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| Immuno-dot blot/Western blot | 5nm-20nm | Protein conjugated gold nanoparticles (antibodies, streptavidin etc) | Colorimetric straightforward detection (no equipment required) Generates a permanent label |
Immunoblotting protocol for gold conjugates |
| Immunohistochemistry (TEM) | 5nm-40nm | Protein conjugated gold nanoparticles (antibodies, streptavidin etc) | High contrast label | - |
| Flow Cytometry | 50nm-400nm | Gold Size Standards | Ideal for standardization of results between runs and experiments when analyzing particles in the 50nm-400nm range. | - |
| Cellular Uptake | 30nm-60nm | Transferrin gold conjugate | Active uptake through endocytosis | - |
| ? | ? | Standard (citrate) | Non-specific cellular uptake | - |
| ? | ? | Cationic gold (available upon request) | High efficiency non-specific cellular uptake | - |
| Darkfield Microscopy | 50nm-100nm | Gold conjugates | - | - |
| Lateral Flow/Dip-Stick Assays | 30nm-80nm | Standard (citrate) | Allows for development of rapid testing kit, point of care assays | Lateral flow immunoassays |
| ? | ? | NHS | ? | ? |
| ? | ? | Carboxyl | ? | ? |
| ? | ? | Amine | ? | ? |
| ? | ? | Streptavidin | ? | ? |
| ? | ? | Protein A | ? | ? |
| ? | ? | Protein G | ? | ? |
| Tumor Targeting | 30nm-80nm | methoxy-PEG | Allows for passive targeting of certain tumors?in vivo Inert material with low non-specific protein binding in serum |
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| Light Microscopy | 5nm-10nm | Gold secondary antibody conjugates | Ability to label tissue sections for both light and electron microscopy.? Alternative to peroxidase and PAP based stains. Sensitivity can be enhanced with?silver enhancement techniques |
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| ELISA | 5nm-30nm | Gold antibody conjugates | Straightforward colorimetric detection |
我怎么洗我的金納米粒子嗎??
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對于大多數應用程序可以使用我們的金納米粒子沒(méi)有任何額外的清洗步驟。 如果你有一個(gè)敏感的應用程序,需要額外的清洗的最佳方式是通過(guò)離心或過(guò)濾。 推薦的離心速度和時(shí)間請參考?
When stored for a long period of time the?gold nanoparticles?might sediment at the bottom of the flask, which is especially true for larger particle sizes. Prior to use, re-suspend the sedimented particles by swirling until a homogenous solution is obtained.
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To maintain optimal performance, and stability of the colloidal gold, care should be taken to use clean storage containers if using other than supplied with the product.
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Characterization of Gold Nanoparticles
Please read our "Introduction to Gold Nanoparticle Characterization" tech note for information on how to analyze gold nanoparticles and their properties.?
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Washing of Gold Nanoparticles
Although it is not always necessary to wash the gold nanoparticles prior to use, some applications might require additional washing procedures. The easiest way to remove possible contaminants in the nanoparticles solution is by centrifugation. Centrifugation force is dependant on size of the gold nanoparticles and should be adjusted according to table I for optimal performance.
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Note:
Since non-functionalized gold nanoparticles are sensitive to salt containing buffers, re-suspension should always be performed in ultra-pure water to prevent irreversible aggregation. Irreversible aggregation is characterized by a clear to bluish solution upon the addition of salt.
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Procedure
- Place aliquot of?colloidal gold?in appropriate centrifuge tube.*
- Centrifuge the gold nanoparticles for 30 minutes using the appropriate G force depending on size of the gold nanoparticles, see Table I.
- Remove supernatant and re-suspend in appropriate volume of ultra-pure water.
- Vortex to re-disperse particles.
*Note: Addition of Tween 20 to a final concentration of 0.025% (w/v) at this step improves performance during centrifugation and can prevent the formation of aggregates. However, Tween 20 binds to the gold surface and can slightly affect the adsorption of other molecules to the gold surface.?
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Table I.?Appropriate G forces for centrifugation of gold nanoparticles. Note that recommended conditions are for a volume of 1ml and centrifugation using a microcentrifuge, except for 5nm gold nanoparticles that requires an ultracentrifuge.
| Size (nm) | Speed (g) | Time (min) |
|---|---|---|
| 5 | 100,000 | 30 |
| 10 | 17,000 | 60 (~50% recovery) |
| 15 | 17,000 | 30 |
| 20 | 6,500 | 30 |
| 30 | 4,500 | 30 |
| 40 | 2,500 | 30 |
| 50 | 2,000 | 30 |
| 60 | 1,125 | 30 |
| 80 | 600 | 30 |
| 100 | 400 | 30 |
| 150 | 180 | 30 |
| 200 | 100 | 30 |
處理?
當存儲在很長(cháng)一段時(shí)間?金納米粒子?可能在燒瓶底部的沉積物,特別是較大的粒子大小。 使用之前,re-suspend沉淀粒子的旋轉,直到齊次解。?
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保持最佳性能,穩定的膠體金,應該小心使用干凈的存儲容器如果使用除了提供產(chǎn)品。?
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金納米粒子的表征
請閱讀我們的“?介紹金納米粒子特性?“信息技術(shù)注意如何分析金納米粒子及其屬性。?
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洗的金納米粒子?
盡管它并不總是必要洗金納米粒子使用之前,一些應用程序可能需要額外的清洗程序。 可能最簡(jiǎn)單的方法去除污染物在納米粒子溶液離心分離。 離心力依賴(lài)于金納米粒子的大小,應根據表調整為最佳性能。?
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注意:?
自non-functionalized金納米粒子對鹽敏感包含緩沖區,re-suspension總是應該在超純水,防止不可逆聚合。 添加鹽是一個(gè)不可逆聚合。
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過(guò)程
- 地方整除的?膠體金?在合適的離心管。*
- 離心機30分鐘的金納米粒子使用適當的G力根據金納米粒子的大小不同,見(jiàn)下表。
- 去除上層清液和re-suspend適當體積的超純水。
- 渦re-disperse粒子。
*注意:添加漸變20的終濃度0.025%(w / v)在離心這一步提高了性能,可以防止骨料的形成。 然而,漸變20金表面結合,可以稍微影響其他分子的吸附金的表面。?
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表我。?適當的G力離心金納米粒子。 注意,推薦條件1毫升的體積和使用微型離心機離心,除了5納米金納米粒子,需要一個(gè)超離心機。
| 大小(nm) | 速度(?g?) | 時(shí)間(分鐘) |
|---|---|---|
| 5 | 100000年 | 30 |
| 10 | 17000年 | 60(~ 50%恢復) |
| 15 | 17000年 | 30 |
| 20 | 6500年 | 30 |
| 30 | 4500年 | 30 |
| 40 | 2500年 | 30 |
| 50 | 2000年 | 30 |
| 60 | 1125年 | 30 |
| 80年 | 600年 | 30 |
| 100年 | 400年 | 30 |
| 150年 | 180年 | 30 |
| 200年 | 100年 | 30 |
我該什么黃金納米顆粒大小選擇對于我的應用程序??
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金納米顆粒的大小和表面化學(xué)是非常依賴(lài)于應用程序使用。 請參閱我們的?黃金納米粒子產(chǎn)品選擇指南?找到最優(yōu)的產(chǎn)品為您的特定應用程序。
我的金納米粒子在存儲的解決方案。 這是正常的嗎??
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的金納米粒子沉降存儲瓶的底部是完全正常的,常用于大尺寸的顆粒,以更大的速度解決。 沉淀不影響粒子的性能。 使用之前,只是漩渦解決適當分散你的金納米粒子和生成一個(gè)同質(zhì)的解決方案。
金納米粒子的保質(zhì)期是什么??
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我們的標準金納米粒子穩定至少1年如果存儲未開(kāi)封和指定的在4攝氏度。 不存儲non-functionalized標準金納米粒子在冰箱里因為這導致不可逆轉的聚合,看到我們嗎?金納米粒子處理和存儲?技術(shù)報告。
金納米粒子溶液變成紫色當我添加包含緩沖鹽。 這是為什么呢??
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由于排斥力的金納米粒子的表面電荷,單個(gè)粒子的能量勢壘必須克服互動(dòng)。 當沒(méi)有(或小)大量的電解質(zhì),如氯化鈉存在,這種能量勢壘過(guò)于強烈的粒子之間的相互作用發(fā)生。 然而,在添加氯化鈉這能量勢壘降低使金納米粒子相互作用和聚合。 這聚合會(huì )導致一個(gè)現象稱(chēng)為表面等離子體耦合,改變光到一個(gè)更高的吸附最大波長(cháng)從而產(chǎn)生的顏色變化的解決方案。
如果我不小心凍結non-functionalized金納米粒子嗎??
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凍結我們的non-functionalized金納米粒子引起不可逆轉的聚合,可以觀(guān)察到明顯的顏色變化的解決方案,見(jiàn)我們?金納米粒子處理和存儲?技術(shù)的更多信息和示例,請注意。
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關(guān)于我們
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- Email: info@wolcavi.com
