Ordering
| Item | Catalog # | Description | Quantity | Price (USD) | ||
|---|---|---|---|---|---|---|
| Plasmid | 104492 | Standard format: Plasmid sent in bacteria as agar stab | 1 | $75 | Add to Cart | |
| AAV1 | 104492-AAV1 | Virus (100 µL at titer ≥ 1×10¹³ vg/mL)and Plasmid.More Information | Add to Cart | |||
| AAV8 | 104492-AAV8 | Virus (100 µL at titer ≥ 1×10¹³ vg/mL)and Plasmid.More Information | Add to Cart | |||
| AAV9 | 104492-AAV9 | Virus (100 µL at titer ≥ 1×10¹³ vg/mL)and Plasmid.More Information | Add to Cart | |||
| AAV Retrograde | 104492-AAVrg | Virus (100 µL at titer ≥ 7×10¹² vg/mL)and Plasmid.More Information | Add to Cart | |||
| AAV PHP.eB | 104492-PHPeB | Virus (100 µL at titer ≥ 1×10¹³ vg/mL)and Plasmid.More Information | Add to Cart | |||
This material is available to academics and nonprofits only.
Backbone
- Vector backboneAAV-Syn-FLEX(Search Vector Database)
- Backbone manufacturerScott Sternson
- Backbone sizew/o insert(bp)4894
- Total vector size (bp)6247
- Vector typeMammalian Expression, AAV, Cre/Lox
Growth in Bacteria
- Bacterial Resistance(s)Ampicillin
- Growth Temperature30°C
- Growth Strain(s)NEB Stable
- Copy numberLow Copy
Gene/Insert
- Gene/Insert namejGCaMP7f
- Alt nameGCaMP3-T302L R303P A317L D380Y
- Alt nameGCaMP3 variant 921
- Alt nameJanelia GCaMP7
- SpeciesR. norvegicus (rat), G. gallus (chicken); A. victoria (jellyfish)
- Insert Size (bp)1353
- PromoterSynapsin
- Tag/ Fusion Protein
- T7 epitope, Xpress tag, 6xHis
Cloning Information
- Cloning methodRestriction Enzyme
- 5′ cloning siteBsmBI(destroyed during cloning)
- 3′ cloning siteBsmBI(destroyed during cloning)
- 5′ sequencing primerACCACGCGAGGCGCGAGATAG (Common Sequencing Primers)
Resource Information
- Terms and Licenses
- UBMTA
- Ancillary Agreement for Plasmids Containing FP Materials
- genOway Notice of RIghts
- Industry Terms
- Not Available to Industry
Information for AAV1 (Catalog # 104492-AAV1)(Back to top)
Purpose
Ready-to-use AAV1 particles produced from pGP-AAV-syn-FLEX-jGCaMP7f-WPRE (#104492). In addition to the viral particles, you will also receive purified pGP-AAV-syn-FLEX-jGCaMP7f-WPRE plasmid DNA.
Syn-driven, Cre-dependent GCaMP7f calcium sensor. These AAV preparations are suitable purity for injection into animals.Delivery
- Volume100 µL
- Titer≥ 1×10¹³ vg/mL
- Pricing$350 USD for preparation of 100 µL virus + $30 USD for plasmid.
- StorageStore at -80℃. Thaw just before use and keep on ice.
- ShipmentViral particles are shipped frozen on dry ice. Plasmid DNA (≥ 200ng) will also be included in the shipment.
Viral Production & Use
- Packaging Plasmidsencode adenoviral helper sequences and AAV rep gene, AAV1 cap gene
- BufferPBS + 0.001% Pluronic F-68
- SerotypeAAV1
- PurificationIodixanol gradient ultracentrifugation
Biosafety
Requestor is responsible for compliance withtheir institution"s biosafety regulations.Lentivirus is generally considered BSL-2. AAV isgenerally considered BSL-1, but may requireBSL-2 handling depending on the insert.Biosafety Guide
Resource Information
- Terms and Licenses
- Ancillary Agreement for Penn Vectors
- Terms of Use for Viral Vectors
- Industry Terms
- Not Available to Industry
Viral Quality Control
- Addgene ensures high quality viral vectors by optimizing and standardizing production protocols and performing rigorous quality control (QC) (see a list of our QC assays). Thespecific QC assays performed varies for each viral lot. To learn which specific QC assays were performed on your lot, please contact us.
- Titer: the exact titer of your sample will be reported on the tube. The titer you see listed on this page is the guaranteed minimum titer. See how titers are measured.
Visit our viral production page for moreinformation.
Addgene Comments
Using FLEX vectors in vivo: LoxP sites in FLEX plasmids are known to recombine during DNA amplification and viral vector production, which may result in a minority of Cre-activated (i.e., "flipped") viral vectors. Addgene has measured this occurs in 0.01-0.03% of viral vectors in our typical production protocol. This can lead to a small number of cells exhibiting Cre-independent transgene expression in vivo. To address this, we recommend titrating to find the optimal AAV dosage required for Cre-dependent transgene expression and function in vivo. This may include reducing the viral vector dosage in order to reduce the likelihood of Cre-independent expression.
Information for AAV8 (Catalog # 104492-AAV8)(Back to top)
Purpose
Ready-to-use AAV8 particles produced from pGP-AAV-syn-FLEX-jGCaMP7f-WPRE (#104492). In addition to the viral particles, you will also receive purified pGP-AAV-syn-FLEX-jGCaMP7f-WPRE plasmid DNA.
Syn-driven, Cre-dependent GCaMP7f calcium sensor. These AAV preparations are suitable purity for injection into animals.Delivery
- Volume100 µL
- Titer≥ 1×10¹³ vg/mL
- Pricing$350 USD for preparation of 100 µL virus + $30 USD for plasmid.
- StorageStore at -80℃. Thaw just before use and keep on ice.
- ShipmentViral particles are shipped frozen on dry ice. Plasmid DNA (≥ 200ng) will also be included in the shipment.
Viral Production & Use
- Packaging Plasmidsencode adenoviral helper sequences and AAV rep gene, AAV8 cap gene
- BufferPBS + 0.001% Pluronic F-68
- SerotypeAAV8
- PurificationIodixanol gradient ultracentrifugation
Biosafety
Requestor is responsible for compliance withtheir institution"s biosafety regulations.Lentivirus is generally considered BSL-2. AAV isgenerally considered BSL-1, but may requireBSL-2 handling depending on the insert.Biosafety Guide
Resource Information
- Terms and Licenses
- Ancillary Agreement for Penn Vectors
- Terms of Use for Viral Vectors
- Industry Terms
- Not Available to Industry
Viral Quality Control
- Addgene ensures high quality viral vectors by optimizing and standardizing production protocols and performing rigorous quality control (QC) (see a list of our QC assays). Thespecific QC assays performed varies for each viral lot. To learn which specific QC assays were performed on your lot, please contact us.
- Titer: the exact titer of your sample will be reported on the tube. The titer you see listed on this page is the guaranteed minimum titer. See how titers are measured.
Visit our viral production page for moreinformation.
Addgene Comments
Using FLEX vectors in vivo: LoxP sites in FLEX plasmids are known to recombine during DNA amplification and viral vector production, which may result in a minority of Cre-activated (i.e., "flipped") viral vectors. Addgene has measured this occurs in 0.01-0.03% of viral vectors in our typical production protocol. This can lead to a small number of cells exhibiting Cre-independent transgene expression in vivo. To address this, we recommend titrating to find the optimal AAV dosage required for Cre-dependent transgene expression and function in vivo. This may include reducing the viral vector dosage in order to reduce the likelihood of Cre-independent expression.
Information for AAV9 (Catalog # 104492-AAV9)(Back to top)
Purpose
Ready-to-use AAV9 particles produced from pGP-AAV-syn-FLEX-jGCaMP7f-WPRE (#104492). In addition to the viral particles, you will also receive purified pGP-AAV-syn-FLEX-jGCaMP7f-WPRE plasmid DNA.
Syn-driven, Cre-dependent GCaMP7f calcium sensor. These AAV preparations are suitable purity for injection into animals.Delivery
- Volume100 µL
- Titer≥ 1×10¹³ vg/mL
- Pricing$350 USD for preparation of 100 µL virus + $30 USD for plasmid.
- StorageStore at -80℃. Thaw just before use and keep on ice.
- ShipmentViral particles are shipped frozen on dry ice. Plasmid DNA (≥ 200ng) will also be included in the shipment.
Viral Production & Use
- Packaging Plasmidsencode adenoviral helper sequences and AAV rep gene, AAV9 cap gene
- BufferPBS + 0.001% Pluronic F-68
- SerotypeAAV9
- PurificationIodixanol gradient ultracentrifugation
Biosafety
Requestor is responsible for compliance withtheir institution"s biosafety regulations.Lentivirus is generally considered BSL-2. AAV isgenerally considered BSL-1, but may requireBSL-2 handling depending on the insert.Biosafety Guide
Resource Information
- Terms and Licenses
- Ancillary Agreement for Penn Vectors
- Terms of Use for Viral Vectors
- Industry Terms
- Not Available to Industry
Viral Quality Control
- Addgene ensures high quality viral vectors by optimizing and standardizing production protocols and performing rigorous quality control (QC) (see a list of our QC assays). Thespecific QC assays performed varies for each viral lot. To learn which specific QC assays were performed on your lot, please contact us.
- Titer: the exact titer of your sample will be reported on the tube. The titer you see listed on this page is the guaranteed minimum titer. See how titers are measured.
Visit our viral production page for moreinformation.
Addgene Comments
Using FLEX vectors in vivo: LoxP sites in FLEX plasmids are known to recombine during DNA amplification and viral vector production, which may result in a minority of Cre-activated (i.e., "flipped") viral vectors. Addgene has measured this occurs in 0.01-0.03% of viral vectors in our typical production protocol. This can lead to a small number of cells exhibiting Cre-independent transgene expression in vivo. To address this, we recommend titrating to find the optimal AAV dosage required for Cre-dependent transgene expression and function in vivo. This may include reducing the viral vector dosage in order to reduce the likelihood of Cre-independent expression.
Information for AAV Retrograde (Catalog # 104492-AAVrg)(Back to top)
Purpose
Ready-to-use AAV Retrograde particles produced from pGP-AAV-syn-FLEX-jGCaMP7f-WPRE (#104492). In addition to the viral particles, you will also receive purified pGP-AAV-syn-FLEX-jGCaMP7f-WPRE plasmid DNA.
Syn-driven, Cre-dependent GCaMP7f calcium sensor. These AAV preparations are suitable purity for injection into animals. These AAV were produced with a retrograde serotype, which permits retrograde access to projection neurons.Delivery
- Volume100 µL
- Titer≥ 7×10¹² vg/mL
- Pricing$350 USD for preparation of 100 µL virus + $30 USD for plasmid.
- StorageStore at -80℃. Thaw just before use and keep on ice.
- ShipmentViral particles are shipped frozen on dry ice. Plasmid DNA (≥ 200ng) will also be included in the shipment.
Viral Production & Use
- Packaging Plasmidsencode adenoviral helper sequences and AAV rep gene, AAV retrograde cap gene from rAAV2-retro helper (plasmid #81070)
- BufferPBS + 0.001% Pluronic F-68 + 200 mM NaCl
- SerotypeAAV retrograde (AAVrg)
- PurificationIodixanol gradient ultracentrifugation
Biosafety
Requestor is responsible for compliance withtheir institution"s biosafety regulations.Lentivirus is generally considered BSL-2. AAV isgenerally considered BSL-1, but may requireBSL-2 handling depending on the insert.Biosafety Guide
Resource Information
- Terms and Licenses
- Terms of Use for Viral Vectors
- Industry Terms
- Not Available to Industry
Viral Quality Control
- Addgene ensures high quality viral vectors by optimizing and standardizing production protocols and performing rigorous quality control (QC) (see a list of our QC assays). Thespecific QC assays performed varies for each viral lot. To learn which specific QC assays were performed on your lot, please contact us.
- Titer: the exact titer of your sample will be reported on the tube. The titer you see listed on this page is the guaranteed minimum titer. See how titers are measured.
Visit our viral production page for moreinformation.
Addgene Comments
Retrograde functionality is dependent on high viral titers. Addgene recommends not diluting your AAV preps prior to use.Using FLEX vectors in vivo: LoxP sites in FLEX plasmids are known to recombine during DNA amplification and viral vector production, which may result in a minority of Cre-activated (i.e., "flipped") viral vectors. Addgene has measured this occurs in 0.01-0.03% of viral vectors in our typical production protocol. This can lead to a small number of cells exhibiting Cre-independent transgene expression in vivo. To address this, we recommend titrating to find the optimal AAV dosage required for Cre-dependent transgene expression and function in vivo. This may include reducing the viral vector dosage in order to reduce the likelihood of Cre-independent expression.
Information for AAV PHP.eB (Catalog # 104492-PHPeB)(Back to top)
Purpose
Ready-to-use AAV PHP.eB particles produced from pGP-AAV-syn-FLEX-jGCaMP7f-WPRE (#104492). In addition to the viral particles, you will also receive purified pGP-AAV-syn-FLEX-jGCaMP7f-WPRE plasmid DNA.
Syn-driven, Cre-dependent GCaMP7f calcium sensor. These AAV were produced with the PHPeB serotype, which permits efficient transduction of the central nervous system. These AAV preparations are suitable purity for injection into animals.Delivery
- Volume100 µL
- Titer≥ 1×10¹³ vg/mL
- Pricing$350 USD for preparation of 100 µL virus + $30 USD for plasmid.
- StorageStore at -80℃. Thaw just before use and keep on ice.
- ShipmentViral particles are shipped frozen on dry ice. Plasmid DNA (≥ 200ng) will also be included in the shipment.
Viral Production & Use
- Packaging Plasmidsencode adenoviral helper sequences and AAV rep gene, PHP.eB cap gene pUCmini-iCAP-PHP.eB (plasmid #103005)
- BufferPBS + 0.001% Pluronic F-68
- SerotypePHPeB (plasmid #103005)
- PurificationIodixanol gradient ultracentrifugation
Biosafety
Requestor is responsible for compliance withtheir institution"s biosafety regulations.Lentivirus is generally considered BSL-2. AAV isgenerally considered BSL-1, but may requireBSL-2 handling depending on the insert.Biosafety Guide
Resource Information
- Terms and Licenses
- Ancillary Agreement for Penn Vectors
- Terms of Use for Viral Vectors
- Industry Terms
- Not Available to Industry
Viral Quality Control
- Addgene ensures high quality viral vectors by optimizing and standardizing production protocols and performing rigorous quality control (QC) (see a list of our QC assays). Thespecific QC assays performed varies for each viral lot. To learn which specific QC assays were performed on your lot, please contact us.
- Titer: the exact titer of your sample will be reported on the tube. The titer you see listed on this page is the guaranteed minimum titer. See how titers are measured.
Visit our viral production page for moreinformation.
Addgene Comments
Using FLEX vectors in vivo: LoxP sites in FLEX plasmids are known to recombine during DNA amplification and viral vector production, which may result in a minority of Cre-activated (i.e., "flipped") viral vectors. Addgene has measured this occurs in 0.01-0.03% of viral vectors in our typical production protocol. This can lead to a small number of cells exhibiting Cre-independent transgene expression in vivo. To address this, we recommend titrating to find the optimal AAV dosage required for Cre-dependent transgene expression and function in vivo. This may include reducing the viral vector dosage in order to reduce the likelihood of Cre-independent expression.
Citation Information: When using the PHP.eB serotype in future publications, please acknowledge Viviana Gradinaru and cite Chan et al., Nat Neurosci, 20(8):1172-1179. Pubmed.ebiomall.com
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TaqDNA聚合酶表达载体的构建与提取纯化
1.背景
重组TaqDNA聚合酶基因及表达载体插入重组TaqDNA聚合酶基因的pET-28a表达质粒，该质粒是由美国Novagen公司推出的能在大肠杆菌中高效表达的一种质粒载体。该质粒在克隆位点前有1个T7启动子，在T7启动子前有1个6×His-Tagcoding序列，是T7的增强子，多克隆位点上有NcoI—Xhol11个酶切位点，载体上还有1个卡那霉素(Kan)的筛选标记。重组的TaqDNA聚合酶基因通过NdeI和Xhol双酶切克隆在表达质粒pET-28a多克隆位点上。
宿主菌为BL21(DE3)。菌株，即宿主菌BL21经噬菌体DE3溶源化后，DE3的lacUV5强启动子及位于其下游的T7RNA聚合酶基因被整合到宿主菌的基因组DNA中。宿主菌在非代谢性乳糖类似物IPTG的诱导作用下能产生大量的T7RNA聚合酶，而T7RNA聚合酶能特异性地识别Pet-28a表达载体中的T7启动子序列，从而高效地表达目的重组蛋白。由于IPTG不会被宿主菌利用，因此向培养液中加入少量的IPTG就能lacUV5强启动子产生持久的诱导作用。
2.材料与方法
1）.TaqDNA聚合酶表达载体构建
Nde1,Sal1双酶切通过pcr引进此双酶切位点taq酶基因，经过T4连接酶连接，将长度为2496bp的taq聚合酶基因，插入同样双酶切的PET28a载体中。
2）.工程菌的转化
通过热击转化的方法，将连接产物转化BL21(DE3)表达菌株，通过菌落pcr,提取质粒酶切鉴定确认taq酶基因已经确实插入PET28a载体，送表达菌株测序验证。
3）.TaqDNA聚合酶基因的诱导表达
取已鉴定的阳性克隆，转接20ml含卡那霉素的LB液体培养基，37℃培养，活化转化细胞。活化后，再取2mL培养液，转接到200mL含卡那霉素的新鲜LB液体培养基，37℃、150r／mim振荡培养3.5h左右，至OD值=1.0时，加入表达诱导剂(IPTG)，至终浓度1．0mmol／L，继续在37℃,150r／min条件下振荡培养，诱导表达，诱导8小时后制备。（转接量、诱导起始时间。诱导用IPTG浓度、诱导时间已经经过系统优化）
4）.10000g/4℃/5min收集菌体，用50ml缓冲液A（20mMTris-HCL,0.2MNaCl,PH8.0）洗涤，10000g/4℃/3min，20ml缓冲液A重悬，4mg/ml终浓度溶菌酶室温处理20分钟，加入DTT使终浓度为1mM,加入吐温-20使体积百分比为0.5%，75℃水浴45分钟，15000g/4℃/20分钟，得到的上清即粗蛋白。上清中加入硫酸铵（4M的母液PH8.0）使其终浓度为1.6M，室温磁力搅拌器搅拌30分钟，12000g/室温/20分钟，用缓冲液B（20mMTris-HCL,100mMKCl,0.5mMEDTA,1mMDTT,0.5℅吐温-20,0.5℅NP-40,50℅甘油，PH8.0）重悬，酶液用缓冲液B4℃透析过夜。
5）.步骤4所得到的酶液即可用于检测实验，为了得到更纯的酶，粗蛋白经中速滤纸过滤后过QsepharoseFF,收集穿透液，再利用工程菌表达的6His作为纯化标记。上清(补足NaCl至终浓度为0.5M)流过已螯合Ni的Ni-NTA柱，该柱先用10倍床体积结合缓冲液（20mMTris-HCL,0.5MNaCl,PH8.0,5mM咪唑）平衡；再用5倍床体积洗涤缓冲溶液（20mMTris-HCL,0.5MNaCl,PH8.0，15mM/30mM咪唑）依次洗涤；最后用洗脱液（20mMTris-HCL,0.1MNaCl,PH8.0,200mM咪唑）洗脱目的蛋白（SDS-PAGE检测蛋白纯度可达到85%以上）。含目的蛋白的洗脱液用sephadexG25脱盐，脱盐缓冲液即步骤4中的缓冲液B。（也可以将洗脱下来的酶液用2M终浓度硫酸铵沉淀后离心，沉淀用酶保存缓冲液溶解，对检测实验没有影响）
反应缓冲液中的镁离子浓度可以根据具体实验需要进行调整
3.检测
纯化后的酶十倍稀释，荧光定量PCR检测活性
SDS-PAGE检测纯度。
4.总结与心得
1）.实验前的准备工作是实验成功的基石，广义的讲：这包括实验大方向的确定、原始文献阅读、基因序列的查找比对、基础工作的准备、每一步实验的分析总结。做好每一步实验的科学分析可以避免后续实验的失败还可以给自己每一小步都成功的感觉，做研发，需要给自己信心。
2）.做好预实验，也就是在实验中做好阳性对照和阴性对照，这有助于我们实验结果的分析。举个例子：做转化时，我们加上质粒转化验证感受态没有问题，加上感受态细胞转化抗生素平板验证感受态没有被污染，这样，无论是什么样的转化结果（包括平板筛选结果）都可以得到很好的分析。
3）.用实验结果证明实验方法的成功与否，在蛋白纯化过程中，特别是在过阴离子柱或者阳离子柱时，凭空想象的离子浓度和PH值是不可取的，只有通过线性的离子浓度和PH值摸索纯化条件、数次重复对比实验，用真实的实验结果来确定所使用的实验条件。
4）.认真原始阅读文献，尽量用最符合生物科学的方法来解决问题，在实验过程中可以参照文献的实验方法，但一切用实验结果说话。
用于包被固相载体的抗原按其来源不同可分为天然抗原、重组抗原和合成多肽抗原三大类。天然抗原可取自动物组织、微生物培养物等，须经提取纯化才能作包被用。如HBsAg可以从携带者的血清中提取，一般的细菌和病毒抗原可以从其培养物中提取，蛋白成份抗原可从富含此抗原的材料中提取等（例如AFP从脐带血或胎肝中提取）。重组抗原是抗原基因在质粒体中表达的蛋白质抗原，多以大肠杆菌或酵母菌为质粒体。重组抗原的优点是除工程菌成份外，其他杂质少，而且无传染性，但纯化技术难度较大。以大肠杆菌为质粒体的重组抗原如不能充分除大肠杆菌成份，用于ELISA试剂盒，在反应中可出现假阳性，因不少受检者受大肠杆菌感染而在血清中存在抗大肠杆菌抗体。重组抗原的另一特点是能用基因工程制备某些无法从天然材料中分离的抗原物质。例如丙型肝炎病毒（HCV）尚不能培养成功，而且丙肝病人血清中HCV抗原含量极微。目前检测抗HCVELISA中所用包被抗原大多为根据HCV的基因克隆表达而制备的重组抗原。在传染病诊断中，不少重组抗原如HBsAg、HBeAg和HIV抗原等均在ELISA中取得应用。合成多肽抗原是根据蛋白质抗原分子的某一抗原决定簇的氨基酸序列人工合成的多肽片段。多肽抗原一般只含有一个抗原决定簇，纯度高，特异性也高，但由于分子量太小，往往难于直接吸附于固相上。多肽抗原的包被一般需先使其与无关蛋白质如牛血清白蛋白质（BSA）等偶联，借助于偶联物与固相载体的吸附，间接地结合到固相载体表面。应用多肽抗原的另一注意点为他仅能检测与其相应的抗体。一种蛋白质抗原往往含有多个不同的能引起抗体产生的决定簇，因此在受检血清中的其他抗体就不能与该多肽抗原发生反应。另外，某些微生物发生变异时往往发生抗原结构变化，在这种情况下，用个别多肽抗原进行包被可引起其他抗体的漏检。 （二）ELISA试剂盒包被用抗体 包被固相载体的抗体应具有高亲和力和高特异性，可取材于抗血清或含单克隆抗体的腹水或培养液。如免疫用抗原中含有杂质（即便是极微量的），在抗血清中将出现杂抗体，必须除去（可用吸收法）后才能用于ELISA，以保证试验的特异性。抗血清不能直接用于包被，应先提取IgG，通常采用硫酸铵盐析和Sephadex凝胶过滤法。一般经硫酸铵盐析粗提的IgG已可用于包被，高度纯化的IgG性质不稳定。如需用高亲和力的抗体包被以提高试验的敏感性，则可采用亲和层析法以除去抗血清中含量较多的非特异性IgG。腹水中单抗的浓度较高，特异性亦较强，因此不需要作吸收和亲和层析处理，一般可将腹水作适当稀释后直接包被，必要时也可用纯化的IgG。应用单抗包被时应注意，一种单抗仅针对一种抗原决定簇，在某些情况下，用多种单抗混合包被，可取得更好的效果。
其滤精度0.01~0.1微米属于二十世纪高新技术种利用压差膜离技术滤除水铁锈、泥沙、悬浮物、胶体、细菌、机物等害物质并能保留体益些矿物质元素
矿泉水、山泉水产工艺核部件超滤工艺水收率高达95%并且便实现冲洗与反冲洗易堵塞使用寿命相较
超滤净水器需要加电加压仅依靠自水压力进行滤流量使用本低廉较适合家庭饮用水全面净化未饮用水净化超滤技术主并结合其滤材料达较宽处理范围更全面消除水污染物质
钠滤（NF）
其滤精度介于超滤反渗透间脱盐率比反渗透低种需要加电、加压膜离技术水加收率较低般用于工业纯水制造
——广东震昊机电希望您帮助
（一）包被用抗原
用于包被固相载体的抗原按其来源不同可分为天然抗原、重组抗原和合成多肽抗原三大类。天然抗原可取自动物组织、微生物培养物等，须经提取纯化才能作包被用。如HBsAg可以从携带者的血清中提取，一般的细菌和病毒抗原可以从其培养物中提取，蛋白成份抗原可从富含此抗原的材料中提取等（例如AFP从脐带血或胎肝中提取）。重组抗原是抗原基因在质粒体中表达的蛋白质抗原，多以大肠杆菌或酵母菌为质粒体。重组抗原的优点是除工程菌成份外，其他杂质少，而且无传染性，但纯化技术难度较大。以大肠杆菌为质粒体的重组抗原如不能充分除大肠杆菌成份，用于ELISA试剂盒，在反应中可出现假阳性，因不少受检者受大肠杆菌感染而在血清中存在抗大肠杆菌抗体。重组抗原的另一特点是能用基因工程制备某些无法从天然材料中分离的抗原物质。例如丙型肝炎病毒（HCV）尚不能培养成功，而且丙肝病人血清中HCV抗原含量极微。目前检测抗HCVELISA中所用包被抗原大多为根据HCV的基因克隆表达而制备的重组抗原。在传染病诊断中，不少重组抗原如HBsAg、HBeAg和HIV抗原等均在ELISA中取得应用。合成多肽抗原是根据蛋白质抗原分子的某一抗原决定簇的氨基酸序列人工合成的多肽片段。多肽抗原一般只含有一个抗原决定簇，纯度高，特异性也高，但由于分子量太小，往往难于直接吸附于固相上。多肽抗原的包被一般需先使其与无关蛋白质如牛血清白蛋白质（BSA）等偶联，借助于偶联物与固相载体的吸附，间接地结合到固相载体表面。应用多肽抗原的另一注意点为他仅能检测与其相应的抗体。一种蛋白质抗原往往含有多个不同的能引起抗体产生的决定簇，因此在受检血清中的其他抗体就不能与该多肽抗原发生反应。另外，某些微生物发生变异时往往发生抗原结构变化，在这种情况下，用个别多肽抗原进行包被可引起其他抗体的漏检。
（二）ELISA试剂盒包被用抗体
包被固相载体的抗体应具有高亲和力和高特异性，可取材于抗血清或含单克隆抗体的腹水或培养液。如免疫用抗原中含有杂质（即便是极微量的），在抗血清中将出现杂抗体，必须除去（可用吸收法）后才能用于ELISA，以保证试验的特异性。抗血清不能直接用于包被，应先提取IgG，通常采用硫酸铵盐析和Sephadex凝胶过滤法。一般经硫酸铵盐析粗提的IgG已可用于包被，高度纯化的IgG性质不稳定。如需用高亲和力的抗体包被以提高试验的敏感性，则可采用亲和层析法以除去抗血清中含量较多的非特异性IgG。腹水中单抗的浓度较高，特异性亦较强，因此不需要作吸收和亲和层析处理，一般可将腹水作适当稀释后直接包被，必要时也可用纯化的IgG。应用单抗包被时应注意，一种单抗仅针对一种抗原决定簇，在某些情况下，用多种单抗混合包被，可取得更好的效果。展开
压力差推力膜滤区超滤膜滤、微孔膜滤逆渗透膜滤三类区根据膜层所能截留粒尺寸或量膜额定孔径范围作区标准则微孔膜(MF)额定孔径范围0.02～10μm;超滤膜(UF)0.001～0.02μm;逆渗透膜(RO)0.0001～0.001μm由知超滤膜适于处理溶液溶质离增浓或采用其离技术所难完胶状悬浮液离超滤膜制膜技术即获预期尺寸窄布微孔技术极其重要孔控制素较根据制膜溶液种类浓度、蒸发及凝聚条件等同同孔径及孔径布超滤膜超滤膜般高离膜用作超滤膜高材料主要纤维素衍物、聚砜、聚丙烯腈、聚酰胺及聚碳酸酯等超滤膜做平面膜、卷式膜、管式膜或空纤维膜等形式广泛用于医药工业、食品工业、环境工程等
我都知道筛用筛东西能细物体放行较截留您听说能筛筛?超膜--种超级筛能尺寸等筛!底超滤膜呢?
超滤膜种具超级筛离功能孔膜孔径几纳米几十纳米说根发丝1‰!膜侧施适压力能筛于孔径溶质离量于500道尔顿、粒径于2～20纳米颗粒超滤膜结构称非称前者各向同性没皮层所向孔隙都属于深层滤;者具较致密表层指状结构主底层表层厚度0.1微米或更并具排列序微孔底层厚度200～250微米属于表层滤工业使用超滤膜般非称膜超滤膜膜材料主要纤维素及其衍物、聚碳酸酯、聚氯乙烯、聚偏氟乙烯、聚砜、聚丙烯腈、聚酰胺、聚砜酰胺、磺化聚砜、交链聚乙烯醇、改性丙烯酸聚合物等等
超滤膜早发高离膜60代超滤装置实现工业化超滤膜工业应用十广泛已新型化工单元操作用于离、浓缩、纯化物制品、医药制品及食品工业;用于血液处理、废水处理超纯水制备终端处理装置我已功利用超滤膜进行草药浓缩提纯超滤膜随着技术进步其筛选功能必改进加强类社贡献越越
