Highlights
- Easy Handling: Bypass chloroform, phase separation and precipitation steps.
- NGS-Ready: Ultra-pure RNA without phenol carryover. No DNA contamination (DNase I included).
- Non-Biased: Complete RNA recovery without miRNA loss.
Description
| Compatibility | TRIzol®, RNAzol®, QIAzol®, TriPure™, TriSure™ and all other acid-guanidinium-phenol based solutions can be used in place of TRI Reagent®. |
|---|---|
| Equipment | Microcentrifuge, vortex, magstand |
| Sample Inactivation | TRI Reagent® (provided with R2101, R2103, and R2105) inhibits RNase activity and inactivates viruses and other infectious agents. |
| Sample Source | Any sample stored and preserved in TRI Reagent®, TRIzol® or similar (animal cells, tissue, bacteria, yeast, fecal, biological fluids, and in vitro processed RNA (e.g., transcription products, DNase-treated or labeled RNA)). |
| Size Range | Total RNA ≥ 17 nt |
| Yield | 10 µg RNA (binding capacity), ≥ 30 µl (elution volume) |
Q1: Is Direct-zol suitable for very small numbers of cells?
Yes, the Direct-zol MicroPrep (#R2060) is designed and capable of purifying RNA down to single cell inputs (picogram amounts). A sensitive quantification method is needed (e.g. Qubit, qPCR, etc.)
Q2: Is DNase I available for individual purchase?
All kit components are available for purchase separately.
Q3: How to store DNase-I following resuspension?
Lyophilized DNase I is stable at room temperature. Once resuspended, store frozen aliquots. Minimize freeze thaw cycles as much as possible. Freeze thaw will lower DNase activity.
Q4: Is the DNase-I treatment necessary?
If the downstream application requires DNA-free RNA, we recommend performing the DNase I treatment.
Q5: Is the kit compatible with samples stored in DNA/RNA Shield?
Yes, bring samples homogenized and stored in DNA/RNA Shield to room temperature (20-30ºC). Add 3 volume of TRIzol/TRI Reagent and mix well. Proceed with RNA Purification.
Q6: Is it possible to extract proteins with the Direct-zol RNA kits?
Yes, proteins can be Acetone Precipitated post RNA binding step. Please request supplementary protocol from Zymo Research Technical Support.
Q7: Can samples be stored in TRIzol/TRI Reagent prior to processing?
Yes, samples in TRIzol/TRI Reagent or similar are stable overnight at room temperature and can be stored frozen (-80C). Be sure to lyse and homogenize the sample well prior to freezing. Bring the sample to room temperature prior to RNA Purification.
Q8: Is it possible to isolate DNA with the Direct-zol RNA kits?
Direct-zol DNA/RNA (D2080) kits can isolate DNA from TRIzol
Q9: Is the RNA suitable for Next-Gen sequencing or other sensitive downstream applications?
Yes, the RNA is high quality (A260/A280 >1.8, A260/A230 >1.8) and suitable for any downstream application, including NGS, RT-PCR, hybridization, etc.
Q10: Which phenol-based reagents are compatible with Direct-zol?
The Direct-zol kits are compatible with TRI Reagent, TRIzol, Qiazol, RNAzol, TriPure, TriSure, etc., and any other acid-guanidinium phenol-based reagents.
Q11: What is the difference between the Direct-zol RNA and Quick-RNA kits?
Direct-zol is for samples stored/collected into TRIzol/similar reagents. Quick-RNA is for all other samples.
Q12: What is the difference between the Direct-zol RNA MiniPrep and the Direct-zol RNA MiniPrep Plus?
Both kits function the same, the only difference is the RNA binding capacity of the column provided with the kit.
Q13: I ran out of RNA Wash Buffer. Can I use something else?
Yes, use 80% ethanol as a substitute. RNA Wash Buffer is also sold separately.
“Before I discovered this kit, I was isolating RNA the old school way with chloroform and it would take half the day to finish the protocol. The Direct-zol RNA Miniprep kit is AWESOME!It took hardly any time, the protocol was so easy, and my RNA quality was SO much better. Honestly, this kit revolutionized my life at the bench.”
-A. Newhart (The Wistar Institute)
“Simple protocol and yielded good quality of RNA. Only one kit working for all type of tissue, cell and especially biological fluids.”
-Mohan K. (University of Illinois, Chicago)
“Previously I used a protocol that took 3 hours, now I can have my RNA in 20 minutes. What is not to like about that? Just one column and two buffers, I love it.”
-Arjan V. (Indiana University)
Read More| Cat # | Name | Size | Price | |
|---|---|---|---|---|
| D4100-2-3 | MagBinding Beads | 3 ml | $66.00 | |
| D4100-2-12 | MagBinding Beads | 12 ml | $125.00 | |
| W1001-30 | DNase/RNase-Free Water | 30 ml | $22.00 | |
| R1060-2-100 | RNA Prep Buffer | 100 ml | $122.00 | |
| R1060-2-25 | RNA Prep Buffer | 25 ml | $40.00 | |
| R2100-1-20 | Direct-zol Binding Buffer Concentrate | 20 ml | $84.00 | |
| R2100-2-200 | Direct-zol MagBead PreWash | 200 ml | $174.00 | |
| R2130-1-120 | MagBead DNA/RNA Wash 1 | 120 ml | $198.00 | |
| R2130-1-30 | MagBead DNA/RNA Wash 1 | 30 ml | $63.00 | |
| R2130-2-20 | MagBead DNA/RNA Wash 2 | 20 ml | $54.00 | |
| R2130-2-80 | MagBead DNA/RNA Wash 2 | 80 ml | $171.00 | |
| C2002 | Collection Plate | 2 Plates | $22.00 | |
| C2003 | Elution Plate | 2 Plates | $19.00 | |
| C2007-8 | 96-Well Plate Cover Foil | 8 Foils | $18.00 | |
| R2050-1-200 | TRI Reagent | 200 ml | $219.00 | |
| E1010-1-4 | DNA Digestion Buffer | 4 mL | $15.00 | |
| E1010 | DNase I Set | 250 U | $56.00 |
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促进生长
预防疾病
因此所有羊抗兔可以结合兔子的同种异型抗体，也就是说只要是兔子产生的抗体，羊抗兔（也叫抗抗体或者2抗）都可与之结合。
在试验中，二抗经常被做了手脚，打上了标记，比如辣根过氧化物酶，经常被用在ELISA，WESTERN等经典的生物学试验中。
1.若都为正常，那么并未有太大问题，只要定期复查就可以了。
2.如果TGAb、TMAb、甲状腺功能均高，那么就是桥本氏甲状腺炎并甲亢，需要抗甲亢治疗。
3.如果TGAb、TMAb高，甲状腺功能下降，那么就是桥本氏甲状腺炎并甲减，需要进行甲减治疗。
4.如果TGAb、TMAb高，甲状腺功能正常，那么就是桥本氏甲状腺炎，无需特殊治疗，只要定期复查甲状腺功能就可能以了，不过这种情况有可能以后演变成甲减或者甲亢。
1.若都为正常，那么并未有太大问题，只要定期复查就可以了。
2.如果TGAb、TMAb、甲状腺功能均高，那么就是桥本氏甲状腺炎并甲亢，需要抗甲亢治疗。
3.如果TGAb、TMAb高，甲状腺功能下降，那么就是桥本氏甲状腺炎并甲减，需要进行甲减治疗。
4.如果TGAb、TMAb高，甲状腺功能正常，那么就是桥本氏甲状腺炎，无需特殊治疗，只要定期复查甲状腺功能就可能以了，不过这种情况有可能以后演变成甲减或者甲亢。
请问这个图是怎么做出来的？
辣根过氧化物酶+DAB显色和免疫荧光染色可以同时在一张脑片里面做吗？
具体是先DAB显色还是先染荧光一抗二抗呢，具体实验步骤是怎样的呢？
第一张图是往血管里打入辣根过氧化物酶，后DAB显色，DIC成像
第二张图是萤光图
目前大量制备单抗的方法主要有两大系统，一是动物体内生产法，这是国内外实验室所广泛采用；另一是体外培养法。
（1）动物体内生产单抗的方法
迄今为止，通常情况下均采用动物体内生产单抗的方法，鉴于绝大多数动物用杂交瘤均由BALB/c小鼠的骨髓瘤细胞与同品系的脾细胞融合而得，因此使用的动物当然首选BALB/c小鼠。本方法即将杂交瘤细胞接种于小鼠腹腔内，在小鼠腹腔内生长杂交瘤，并产生腹水，因而可得到大量的腹水单抗且抗体浓度很高。可见该法操作简便、经济，不过，腹水中常混有小鼠的各种杂蛋白（包括Ig），因此在很多情况下要提纯后才能使用，而且还有污染动物病毒的危险，故而最好用SPF级小鼠。
（2）体外培养生产单抗的方法
总体上讲，杂交瘤细胞系并不是严格的贴壁依赖细胞（anchoragedependentcell，ADC），因此既可以进行单层细胞培养，又可以进行悬浮培养。杂交瘤细胞的单层细胞培养法是各个实验室最常用的手段，即将杂交瘤细胞加入培养瓶中，以含10-15%小牛血清的培养基培养，细胞浓度以1×106-2×106/ml为佳，然后收集培养上清，其中单抗含量约10-50ug/ml。显然，这种方法制备的单抗量极为有限，无疑是不适用于单抗的大规模生产。要想在体外大量制备单抗，就必须进行杂交瘤细胞的大量（高密度）培养。单位体积内细胞数量越多，细胞存活时间越长，单抗的浓度就越高，产量就越大。
