EasySep™ 死细胞去除 (Annexin V) 试剂盒

凋亡细胞（Annexin V）的免疫磁珠去除

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产品号 #17899_C

凋亡细胞（Annexin V）的免疫磁珠去除

产品优势

快捷、操作简单
无需分离柱
兼容EasySep™、“The Big Easy”和EasyEights™磁极

产品组分包括

EasySep™死细胞去除（Annexin V）试剂盒（产品号 #17899）

EasySep™死细胞去除（Annexin V）抗体复合物, 0.5 mL
EasySep™生物素分选抗体复合物，1mL
EasySep™Dextran RapidSpheres™50103磁珠，1 mL

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总览

使用 EasySep™ 死细胞去除 (Annexin V) 试剂盒，通过免疫磁性负选，可高效去除细胞培养或组织制备样本中的凋亡细胞 (Annexin V+)。EasySep™结合了单克隆抗体的特异性和无柱磁性系统的简便性，迄今已广泛应用于发表的研究中超过20年。

EasySep™ 操作流程简单，使用识别 Annexin V的抗体复合物和磁珠标记细胞，再通过 EasySep™ 磁极进行无柱分选，只需倾倒或吸取未标记细胞，即可将标记细胞与未标记细胞分离，Annexin V+ 细胞则保留在管中。经过磁性细胞分离后，即可获得所需的细胞用于下游应用。在细胞凋亡过程中，Annexin V 会与细胞膜外层磷脂酰丝氨酸结合。

了解更多关于免疫磁性 EasySep™ 技术的工作原理。探索更多优化您实验流程的产品，包括培养基、补充剂、抗体等。

Data Figures

Figure 1. Typical Profile for Dead Cell Removal from Human PMNCs Using EasySep™ Dead Cell Removal (Annexin V) Kit

Starting with human polymorphonuclear cells (PMNCs) cultured overnight, the live cell content (AnnexinV-/PI-) of the enriched fraction is typically 69.7± 12.5% (mean ± SD), using the purple EasySep™ Magnet. In the above example, the percentages of live cells in the start and final enriched fractions are 12.8% and 74.9%, respectively.

Figure 2. Typical Profile for Dead Cell Removal from Mouse Splenocytes Using EasySep™ Dead Cell Removal (Annexin V) Kit

Starting with 24- to 48-hour-old mouse splenocytes, the live cell content of the enriched fraction is typically 79.8 ± 11.4% (mean ± SD), using the purple EasySep™ Magnet. In the above example, the percentages of live cells in the start and final enriched fractions are 78.1% and 93.4%, respectively.

Protocols and Documentation

Find supporting information and directions for use in the Product Information Sheet or explore additional protocols below.

Educational Materials (6)

Inhibiting DNA methylation and RNA editing upregulates immunogenic RNA to transform the tumor microenvironment and prolong survival in ovarian cancer. S. Gomez et al. Journal for immunotherapy of cancer 2022 nov

Abstract

BACKGROUND Novel therapies are urgently needed for ovarian cancer (OC), the fifth deadliest cancer in women. Preclinical work has shown that DNA methyltransferase inhibitors (DNMTis) can reverse the immunosuppressive tumor microenvironment in OC. Inhibiting DNA methyltransferases activate transcription of double-stranded (ds)RNA, including transposable elements. These dsRNAs activate sensors in the cytoplasm and trigger type I interferon (IFN) signaling, recruiting host immune cells to kill the tumor cells. Adenosine deaminase 1 (ADAR1) is induced by IFN signaling and edits mammalian dsRNA with an A-to-I nucleotide change, which is read as an A-to-G change in sequencing data. These edited dsRNAs cannot be sensed by dsRNA sensors, and thus ADAR1 inhibits the type I IFN response in a negative feedback loop. We hypothesized that decreasing ADAR1 editing would enhance the DNMTi-induced immune response. METHODS Human OC cell lines were treated in vitro with DNMTi and then RNA-sequenced to measure RNA editing. Adar1 was stably knocked down in ID8 Trp53-/- mouse OC cells. Control cells (shGFP) or shAdar1 cells were tested with mock or DNMTi treatment. Tumor-infiltrating immune cells were immunophenotyped using flow cytometry and cell culture supernatants were analyzed for secreted chemokines/cytokines. Mice were injected with syngeneic shAdar1 ID8 Trp53-/- cells and treated with tetrahydrouridine/DNMTi while given anti-interferon alpha and beta receptor 1, anti-CD8, or anti-NK1.1 antibodies every 3 days. RESULTS We show that ADAR1 edits transposable elements in human OC cell lines after DNMTi treatment in vitro. Combining ADAR1 knockdown with DNMTi significantly increases pro-inflammatory cytokine/chemokine production and sensitivity to IFN-$\beta$ compared with either perturbation alone. Furthermore, DNMTi treatment and Adar1 loss reduces tumor burden and prolongs survival in an immunocompetent mouse model of OC. Combining Adar1 loss and DNMTi elicited the most robust antitumor response and transformed the immune microenvironment with increased recruitment and activation of CD8+ T cells. CONCLUSION In summary, we showed that the survival benefit from DNMTi plus ADAR1 inhibition is dependent on type I IFN signaling. Thus, epigenetically inducing transposable element transcription combined with inhibition of RNA editing is a novel therapeutic strategy to reverse immune evasion in OC, a disease that does not respond to current immunotherapies.

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Single-cell profiling of human dura and meningioma reveals cellular meningeal landscape and insights into meningioma immune response. A. Z. Wang et al. Genome medicine 2022 may

Abstract

BACKGROUND Recent investigations of the meninges have highlighted the importance of the dura layer in central nervous system immune surveillance beyond a purely structural role. However, our understanding of the meninges largely stems from the use of pre-clinical models rather than human samples. METHODS Single-cell RNA sequencing of seven non-tumor-associated human dura samples and six primary meningioma tumor samples (4 matched and 2 non-matched) was performed. Cell type identities, gene expression profiles, and T cell receptor expression were analyzed. Copy number variant (CNV) analysis was performed to identify putative tumor cells and analyze intratumoral CNV heterogeneity. Immunohistochemistry and imaging mass cytometry was performed on selected samples to validate protein expression and reveal spatial localization of select protein markers. RESULTS In this study, we use single-cell RNA sequencing to perform the first characterization of both non-tumor-associated human dura and primary meningioma samples. First, we reveal a complex immune microenvironment in human dura that is transcriptionally distinct from that of meningioma. In addition, we characterize a functionally diverse and heterogenous landscape of non-immune cells including endothelial cells and fibroblasts. Through imaging mass cytometry, we highlight the spatial relationship among immune cell types and vasculature in non-tumor-associated dura. Utilizing T cell receptor sequencing, we show significant TCR overlap between matched dura and meningioma samples. Finally, we report copy number variant heterogeneity within our meningioma samples. CONCLUSIONS Our comprehensive investigation of both the immune and non-immune cellular landscapes of human dura and meningioma at single-cell resolution builds upon previously published data in murine models and provides new insight into previously uncharacterized roles of human dura.

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Cancer cell-intrinsic resistance to BiTE therapy is mediated by loss of CD58 costimulation and modulation of the extrinsic apoptotic pathway. Y. Shen et al. Journal for immunotherapy of cancer 2022 mar

Abstract

BACKGROUND Bispecific T-cell engager (BiTE) molecules induce redirected lysis of cancer cells by T cells and are an emerging modality for solid tumor immunotherapy. While signs of clinical activity have been demonstrated, efficacy of T-cell engagers (TCEs) in solid tumors settings, molecular determinants of response, and underlying mechanisms of resistance to BiTE therapy require more investigation. METHODS To uncover cancer cell-intrinsic genetic modifiers of TCE-mediated cytotoxicity, we performed genome-wide CRISPR (Clustered Regularly Interspaced Short Palindromic Repeats) loss-of-function and CRISPRa (CRISPR activation) gain-of-function screens using TCEs against two distinct tumor-associated antigens (TAAs). By using in vitro T-cell cytotoxicity assays and in vivo efficacy studies, we validated the roles of two common pathways identified in our screen, T-cell costimulation pathway and apoptosis pathway, as key modifiers of BiTE activity. RESULTS Our genetic screens uncovered TAAs-independent cancer cell-intrinsic genes with functions in autophagy, T-cell costimulation, the apoptosis pathway, chromatin remodeling, and cytokine signaling that altered responsiveness to BiTE-mediated killing. Notably, loss of CD58 (the ligand of the CD2 T-cell costimulatory receptor), a gene frequently altered in cancer, led to decreased TCE-mediated cytotoxicity, T-cell activation and antitumor efficacy in vitro and in vivo. Moreover, the effects of CD58 loss were synergistically compounded by concurrent loss of CD80/CD86 (ligands for the CD28 T-cell costimulatory receptor), whereas joint CD2 and CD28 costimulation additively enhanced TCE-mediated killing, indicating non-redundant costimulatory mechanisms between the two pathways. Additionally, loss of CFLAR (Caspase-8 and FADD Like Apoptosis Regulator), BCL2L1, and BID (BH3 Interacting Domain Death Agonist) induced profound changes in sensitivity to TCEs, indicating that key regulators of apoptosis, which are frequently altered in cancer, impact tumor responsiveness to BiTE therapy. CONCLUSIONS This study demonstrates that genetic alterations central to carcinogenesis and commonly detected in cancer samples lead to significant modulation of BiTE antitumor activity in vitro and in vivo, findings with relevance for a better understanding of patient responses to BiTE therapy and novel combinations that enhance TCE efficacy.

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Species	Human, Mouse, Non-Human Primate, Other, Rat
Magnet Compatibility	• EasySep™ Magnet (Catalog #18000), or • “The Big Easy” EasySep™ Magnet (Catalog #18001), or • EasyEights™ EasySep™ Magnet (Catalog #18103)
Sample Source	Cord Blood, Leukapheresis, Lung, Lymph Node, Other, Spleen
Selection Method	Depletion

EasySep™ 死细胞去除 (Annexin V) 试剂盒

产品号 #（选择产品）

产品号 #17899_C

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总览

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EasySep™ 死细胞去除 (Annexin V) 试剂盒

产品号 #（选择产品）

产品号 #17899_C

产品优势

产品组分包括

总览

Data Figures

Protocols and Documentation

Applications

Resources and Publications

Educational Materials (6)

Publications (8)

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