RosetteSep™人NK细胞富集抗体混合物

免疫密度负选试剂混合物

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产品号 #（选择产品）

产品号 #15025_C

免疫密度负选试剂混合物

产品优势

快捷、操作简单
不需要特殊设备或额外培训
分选得到的细胞不带标记
可与SepMate™联合使用，实现一致的高通量样本处理

产品组分包括

RosetteSep™人NK细胞富集抗体混合物（产品号 #15025）

RosetteSep™人NK细胞富集抗体混合物，2mL

RosetteSep™人NK细胞富集抗体混合物（产品号 #15065）

RosetteSep™人NK细胞富集抗体混合物，5x2mL

立即询价

总览

RosetteSep™人NK细胞富集抗体混合物通过负选从全血分离NK细胞。四聚体抗体复合物可识别非NK细胞和红细胞（RBC），从而靶向去除非目的细胞。使用密度梯度离心液（如RosetteSep™ DM-L（产品号 #15705）或Lymphoprep™（产品号 #18060）离心后，非目的细胞会与红细胞一起沉淀。纯化的NK细胞为血浆和密度梯度离心液的交界界面中高度富集的细胞。

实验数据

Figure 1. FACS Histogram Results Using RosetteSep™ Human NK Cell Enrichment Cocktail

Starting with fresh peripheral blood the CD56+ cell content of the enriched fraction typically ranges from 80 - 98%. *Note: Red blood cells were removed by lysis prior to flow cytometry.

产品说明书及文档

请在《产品说明书》中查找相关支持信息和使用说明，或浏览下方更多实验方案。

应用领域

本产品专为以下研究领域设计，适用于工作流程中的高亮阶段。探索这些工作流程，了解更多我们为各研究领域提供的其他配套产品。

常见问题

What is RosetteSep™?

RosetteSep™ is a rapid cell separation procedure for the isolation of purified cells directly from whole blood, without columns or magnets.

How does RosetteSep™ work?

The antibody cocktail crosslinks unwanted cells to red blood cells (RBCs), forming rosettes. The unwanted cells then pellet with the free RBCs when centrifuged over a density centrifugation medium (e.g. Ficoll-Paque™ PLUS, Lymphoprep™).

What factors affect cell recovery?

The temperature of the reagents can affect cell recovery. All reagents should be at room temperature (sample, density centrifugation medium, PBS, centrifuge) before performing the isolations. Layering can also affect recovery so be sure to carefully layer the sample to avoid mixing with the density centrifugation medium as much as possible. Be sure to collect the entire enriched culture without disturbing the RBC pellet. A small amount of density centrifugation medium can be collected without worry.

Which cell samples can RosetteSep™ be used with?

RosetteSep™ can be used with leukapheresis samples, bone marrow or buffy coat, as long as: the concentration of cells does not exceed 5 x 10⁷ per mL (can dilute if necessary); and there are at least 100 RBCs for every nucleated cell (RBCs can be added if necessary).

Can RosetteSep™ be used with previously frozen or cultured cells?

Yes. Cells should be re-suspended at 2 - 5 x 10⁷ cells / mL in PBS + 2% FBS. Fresh whole blood should be added at 250 µL per mL of sample, as a source of red cells.

Can RosetteSep™ be used to enrich progenitors from cord blood?

Yes. Sometimes cord blood contains immature nucleated red cells that have a lower density than mature RBCs. These immature red cells do not pellet over Ficoll™, which can lead to a higher RBC contamination than peripheral blood separations.

Does RosetteSep™ work with mouse cells?

No, but we have developed EasySep™, a magnetic-based cell isolation system which works with mouse and other non-human species.

Which anticoagulant should be used with RosetteSep™?

Peripheral blood should be collected in heparinized Vacutainers. Cord blood should be collected in ACD.

Should the anticoagulant be washed off before using RosetteSep™?

No, the antibody cocktail can be added directly to the sample.

文献 (64)

The intracellular granzyme B inhibitor, proteinase inhibitor 9, is up-regulated during accessory cell maturation and effector cell degranulation, and its overexpression enhances CTL potency. Hirst CE et al. Journal of immunology (Baltimore,Md. : 1950) 2003 JAN

Abstract

Granzyme B (grB) is a serine proteinase released by cytotoxic lymphocytes (CLs) to kill abnormal cells. GrB-mediated apoptotic pathways are conserved in nucleated cells; hence,CLs require mechanisms to protect against ectopic or misdirected grB. The nucleocytoplasmic serpin,proteinase inhibitor 9 (PI-9),is a potent inhibitor of grB that protects cells from grB-mediated apoptosis in model systems. Here we show that PI-9 is present in CD4(+) cells,CD8(+) T cells,NK cells,and at lower levels in B cells and myeloid cells. PI-9 is up-regulated in response to grB production and degranulation,and associates with grB-containing granules in activated CTLs and NK cells. Intracellular complexes of PI-9 and grB are evident in NK cells,and overexpression of PI-9 enhances CTL potency,suggesting that cytoplasmic grB,which may threaten CL viability,is rapidly inactivated by PI-9. Because dendritic cells (DCs) acquire characteristics similar to those of target cells to activate naive CD8(+) T cells and therefore may also require protection against grB,we investigated the expression of PI-9 in DCs. PI-9 is evident in thymic DCs (CD3(-),CD4(+),CD8(-),CD45(+)),tonsillar DCs,and DC subsets purified from peripheral blood (CD16(+) monocytes and CD123(+) plasmacytoid DCs). Furthermore,PI-9 is expressed in monocyte-derived DCs and is up-regulated upon TNF-alpha-induced maturation of monocyte-derived DCs. In conclusion,the presence and subcellular localization of PI-9 in leukocytes and DCs are consistent with a protective role against ectopic or misdirected grB during an immune response.

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DNA methylation maintains allele-specific KIR gene expression in human natural killer cells. Chan H-W et al. The Journal of experimental medicine 2003 JAN

Abstract

Killer immunoglobulin-like receptors (KIR) bind self-major histocompatibility complex class I molecules,allowing natural killer (NK) cells to recognize aberrant cells that have down-regulated class I. NK cells express variable numbers and combinations of highly homologous clonally restricted KIR genes,but uniformly express KIR2DL4. We show that NK clones express both 2DL4 alleles and either one or both alleles of the clonally restricted KIR 3DL1 and 3DL2 genes. Despite allele-independent expression,3DL1 alleles differed in the core promoter by only one or two nucleotides. Allele-specific 3DL1 gene expression correlated with promoter and 5' gene DNA hypomethylation in NK cells in vitro and in vivo. The DNA methylase inhibitor,5-aza-2'-deoxycytidine,induced KIR DNA hypomethylation and heterogeneous expression of multiple KIR genes. Thus,NK cells use DNA methylation to maintain clonally restricted expression of highly homologous KIR genes and alleles.

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KIR2DL4 (CD158d) genotype influences expression and function in NK cells. Goodridge JP et al. Journal of immunology (Baltimore,Md. : 1950) 2003 AUG

Abstract

The expression and function of the NK cell receptor KIR2DL4 are controversial. Two common alleles of the transmembrane domain of KIR2DL4 exist. The 10A allele with 10 adenines at the end of the transmembrane exon encodes a full length receptor,whereas the 9A allele has only 9 adenines resulting in a frame shift which in turn generates a stop codon early in the first cytoplasmic exon. The possibility that the 10A and 9A alleles might result in differences in expression and function of KIR2DL4 was explored using mAbs to KIR2DL4. Transfection experiments with cDNA from the 10A and 9A alleles revealed significant membrane expression only with the protein encoded by the 10A allele. Analysis of peripheral blood NK cells demonstrated that only in subjects with at least one 10A allele was cell surface expression of KIR2DL4 detectable,and then only on the minor CD56(bright) NK cell subset. The major CD56(dim) NK cell subset did not cell surface express KIR2DL4 but,interestingly,did so after in vitro culture. Functional analysis using cultured NK cells in redirected lysis assays demonstrated that KIR2DL4 is an activating receptor for NK cells with at least one 10A allele. No significant activity was detected for NK cells generated from subjects homozygous for the 9A allele. These data show that genotype influences cell surface expression and function of KIR2DL4 which may account for reported differences in KIR2DL4 expression and function.

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物种	人
样本来源	全血, 白膜层
Selection Method	Negative

RosetteSep™人NK细胞富集抗体混合物

产品号 #15065

产品号 #15025

产品号 #（选择产品）

产品号 #15025_C

产品优势

产品组分包括

总览

实验数据

产品说明书及文档

应用领域

相关材料与文献

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RosetteSep™人NK细胞富集抗体混合物

产品号 #15065

产品号 #15025

产品号 #（选择产品）

产品号 #15025_C

产品优势

产品组分包括

总览

实验数据

产品说明书及文档

应用领域

相关材料与文献

技术资料 (7)

常见问题

What is RosetteSep™?

How does RosetteSep™ work?

What factors affect cell recovery?

Which cell samples can RosetteSep™ be used with?

Can RosetteSep™ be used with previously frozen or cultured cells?

Can RosetteSep™ be used to enrich progenitors from cord blood?

Does RosetteSep™ work with mouse cells?

Which anticoagulant should be used with RosetteSep™?

Should the anticoagulant be washed off before using RosetteSep™?

文献 (64)

Abstract

Abstract

Abstract

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