By leveraging Takara Bio"s expertise in single-cell RNA-seq library construction and the cell isolation and selection capabilities of the ICELL8 platform, the ICELL8 3" DE and ICELL8 3" DE for UMI workflows enable high-throughput preparation of Illumina libraries for differential expression analysis of single cells from up to eight different samples in parallel.
By leveraging Takara Bio"s expertise in single-cell RNA-seq library construction and the cell isolation and selection capabilities of ICELL8 systems, the ICELL8 3" DE and ICELL8 3" DE for UMI workflows enable high-throughput preparation of Illumina libraries for differential expression analysis of single cells from up to eight different samples in parallel.
The ICELL8 Chip and Reagent 3" DE Kit (Cat. # 640164, 640165, 640166) is compatible with the original ICELL8 Single-Cell System. Included in the kit are an ICELL8 3" DE Chip (Cat. # 640143), a 5,184-nanowell SmartChip that contains preprinted oligo(dT) primers, and reagents for cDNA synthesis and Illumina library prep.
Following staining and dilution, sample cells are dispensed into chip nanowells using the ICELL8 MultiSample NanoDispenser (MSND). Wells containing single, viable cells are selectedusing the ICELL8 Imaging System and the CellSelect Software provided with the ICELL8 system. Following the selection of cells for processing and analysis, RT-PCR reagents are dispensed into the corresponding wells, and cDNA synthesis and amplification are performed in-chip in a single run. Following RT-PCR, cDNA products are pooled and purified, and sequencing libraries are prepared using a transposase-based method that enriches for cDNA derived from mRNA 3" ends, followed by a limited-cycle PCR amplification step that incorporates Illumina index sequences.
Sequencing data from the resulting libraries can be traced back to individual cells using the nanowell-specific barcode sequences and can be used to perform differential gene expression analysis by mRNA 3" end-counting. By yielding high-quality, 3"-focused sequencing data from hundreds of cells in parallel, these kits provide an efficient, cost-effective solution for high-throughput differential expression analysis of single cells.
The ICELL8 3" DE for UMI Reagent Kit is used with the ICELL8 3" DE Chip as part of the ICELL8 3" DE with UMI workflow, which enables Illumina sequencing and differential gene expression analysis-via transcript 3" end counting-of single cells isolated on the ICELL8 system.
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其同型为;percp-cy5.5(25ug包装）浓度：0.2mg/ml,
那么如果CD45RA抗体如果加入20ul，那同型抗体应该加入多少ml:
我自己的初步计算是：1ul的3分之1.不知道我算对了吗？应该怎么加才好呢？
手头上，有eBioscience家的同型抗体，为PE标记（mouseIgG1,k），说明书写使用为2.5ug/test。
按照说明书的浓度加入后结果发现，同型对照抗体组比实验组和阴性对照组（不表达CD80）荧光强度要高，很明确我的同型对照加入的剂量多了，但正确的加法是多少？按照质量算吗？请高手指教，非常感谢！！
英 ['aɪsətaɪp]美 ['aɪsəˌtaɪp]
n.
同型动物（或植物），图形文字，象征性图像;同位型;同模;同号模式
网络
同型;同种型;模式标本
双语例句
Study on the Action of Decreasing level of Isotype Cysteine for High Blood Viscosity
降低同型半胱氨酸水平对高黏血症作用的研究
比如你用anti-MHC-II，mouse IgG1-FITC，那同型就用非特异性的mouse IgG1-FITC，同样浓度
同型对照（阴性对照）管：A同型+B同型+C同型
单阳性对照
A检测管：A+B同型+C同型
B检测管：B+A同型+C同型
C检测管：C+A同型+B同型
不存在荧光重叠的两个抗体不需要调节补偿。
样本为PBMC，想用三色标记Treg细胞，分别为CD4-PECY，CD25-Fitc，Foxp3-PE，想知道如何设置荧光补偿，用的是BD的流式仪，1、是设置阴性管一个，单阳性管每种一个，样品管一个么？
荧光素强度依次为PE， PECY5 APC FITC, PE最强，FITC最弱。 所以，建议抗体组合为：
Foxp3-PE，CD4-FITC，CD25-PECy5.5或者CD25-APC。
同时购买三个相应的同型对照抗体。
样品管设置如下：
1. cell + FITC-isotype + Pecy5.5（ or APC）-isotype + PE-isotype
2. Cell + FITC-isotype + CD25-mAb + PE-isotype
3. Cell + PE-mAb + FITC-isotype + Pecy5.5（ or APC）-isotype
4. Cell + CD4-FITC + Foxp3-PE + CD25 -PECy5.5( or APC)。 [这一管的数据可以拿来分析用]。
在充分了解调整电压和补偿原理的基础上，也可以做一些简化。如果不太懂，就按照这个来。其中Foxp3的抗体和同型对照抗体都是在固定破膜以后再加，不是与CD4或者CD25同时加。
