Description
in vivoDNA RNAAdvanced Transfection Reagent can be injected intravenously or by local tissue injection in vivo. Further more, total injection volume is very small, dense tissue can be transfected by microinjection and the time of circulation in vivo is long. Both DNA, siRNA and co-transfection can becarried out.
General Considerations
1>DNA quality requirements
Concentration: 300ng/ul-2μg/ul;
Solution: dissolved in ddH2O or ultra-pure wate;
Endotoxin: removed
2>RNA quality requirements
Concentration: 20 μM, 40 μM, 60 μM, 80 μM
General Protocol for in vivo Transfection
1>Complex preparation:Nucleic acid was directly mixed with transfection reagent according to 1:1 relationship. Thenuse a pipette to blow up 10-15 times to mix.After incubatingat room temperature for 10-15 minutes,complex would be prepared.During the procedure, no liquid residue was ensured on the wall of tube.
2>Intravenous injection or local tissue injection of prepared complex can be proceeded with a syringe or microinjection needle.
3>3 or 5 days afterinjection, efficiency of transfectionwould reach the peak, and expression of target gene could be detected then.
Important Guidelines
1>The ratio of DNA (μg) or 20 μM siRNA (μl) to transfection reagent (μl) should be 1:1.
2>The volume of siRNA used in above table is specific for siRNA of 20 μM. If the concentration of siRNA is 40 μM, the volume of siRNA in above table should be halved. If the concentration of siRNA is 80 μ M, the volume of siRNA in above table should be divided by 4, and so on.
3>When local tissue injection is performed, the amount of the complex should be 5-10 μl/cm2.
4>In co-transfection experiment, the total amount of nucleic acid in above table remains the same, the proportion of various nucleic acids could be adjusted according to experimental requirements, then nucleic acids can be mixed with transfection reagent sufficiently.
5>In the specific experimental operation, the dosage of nucleic acid and transfection reagent can be adjusted according to the "maximum injection volume" in the table above.
Order Information
Product | Catalog | Size |
in vivo DNA RNA Advanced Transfection Reagent | AV500025 | 0.25ml |
in vivo DNA RNA Advanced Transfection Reagent | AV500050 | 0.50 ml |
in vivo DNA RNA Advanced Transfection Reagent | AV500075 | 0.75ml |
in vivo DNA RNA Advanced Transfection Reagent | AV500150 | 1.50 ml |
in vivo DNA RNA Advanced Transfection Reagent | AV500500 | 5.00 ml |
in vivo DNA RNA Advanced Transfection Reagent | AV501000 | 10.00 ml |
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首先要什么有什么的，你得好好考虑一下。
2、看生产地址
根本没有生产地址，我们知道做实验做产品需要很多的仪器、试剂、耗材，没有人相信一间简单的屋子可以生产各种样的试剂盒。
3、看产品包装
没有任何的生产地址、联系方式等信息，这种产品有问题了连个投诉的地方都没有。
4、看公司网站
有些打着国外原装旗号，整个公司网站为英文页面，实际注册IP地址在中国。如果写着国外的地址，让你国外的朋友实地去看一下！
5、做交叉验证
拿对方提供的几个种类的试剂盒，把里面的关键组份相互替换做做实验，如果交叉严重，只能说明是一种原料生产的试剂盒贴了不同的标签。
6、看价格
价格低得离谱，却打着进口大公司原料分装，核算成本，这种低得离谱的价格是连原料都买不起的。
现在国内最差也是用3代试剂，有些地方会用4代试剂。
4代试剂（检查抗原+抗体）——窗口期为4周。因为抗原于3-4周达到复制的峰值，此时通过4代试剂检查，如果感染了HIV，抗原/抗体至少有一个为阳性，如果都是阴就排除了。
3代试剂（只查抗体）——窗口期为6周。
以上为理论分析+临床经验的结果，可以说是99.9%的准确度。
但是目前FDA、CDC和试剂生产商统一达成的共识，也就是针对普通人，最保守的窗口期是3个月。无论什么试剂，3个月都100%排除。
DAS-ELISA试剂盒中抗体有包被抗体和酶标抗体，这两种抗体是一种抗体，前者没有酶标，后者用酶进行了标记。
NCM-ELISA一般有一抗和二抗，这两种抗体不一样，前者（一抗）一般是鼠抗体或兔抗体，没有酶标，相对应的二抗一般是羊抗鼠或羊抗兔抗体，用酶进行标记。也有用马大规模备抗体的。
TDS-ELISA是三抗体ELISA，第三级的抗体是将信号级联放大。
以上三种是比较常见的，还有其他一些试剂盒，可参考免疫学的内容。
ELISA的基础是抗原或抗体的固相化及抗原或抗体的酶标记。结合在固相载体表面的抗原或抗体仍保持其免疫学活性，酶标记的抗原或抗体既保留其免疫学活性，又保留酶的活性。在测定时，受检标本（测定其中的抗体或抗原）与固相载体表面的抗原或抗体起反应。用洗涤的方法使固相载体上形成的抗原抗体复合物与液体中的其他物质分开。再加入酶标记的抗原或抗体，也通过反应而结合在固相载体上。此时固相上的酶量与标本中受检物质的量呈一定的比例。加入酶反应的底物后，底物被酶催化成为有色产物，产物的量与标本中受检物质的量直接相关，故可根据呈色的深浅进行定性或定量分析。由于酶的催化效率很高，间接地放大了免疫反应的结果，使测定方法达到很高的敏感度。
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