产品号 #01711_C
用于人脐带血CD34+和ALDHbr细胞的检测
若您需要咨询产品或有任何技术问题,请通过官方电话 400 885 9050 或邮箱 info.cn@stemcell.com 与我们联系
用于人脐带血CD34+和ALDHbr细胞的检测
ALDHbr试剂盒是针对人类脐带血样本中表达高水平醛脱氢酶(ALDH)的CD34+细胞的鉴定和定量而优化的。本试剂盒包含CD34+细胞检测试剂盒和ALDEFLUOR™试剂盒。CD34+细胞检测试剂盒包含预先选择的荧光染料偶联单克隆抗体,这些抗体已经过优化,可与ALDEFLUOR™试剂盒一起使用,通过流式细胞术检测表达CD34并具有高ALDH活性的细胞。CD34+细胞检测试剂盒包含抗人抗体APC CD34, PE CD45, PE- cyanine5 Glycophorin A/B, 7-AAD活性染料。
需要配备488 nm激光器和635 nm激光器的流式细胞仪,用于检测APC, PE, PE- cyanine5和7-AAD荧光。
Cell Type
Hematopoietic Stem and Progenitor Cells
Species
Human
Application
Flow Cytometry
Area of Interest
Cord Blood Banking, Stem Cell Biology
Figure 1. General ALDHbr Assay Procedure
For a detailed protocol please refer to the ALDHbr Assay Kit Product Information Sheet (PIS)
Figure 2. Gating Strategy to Identify 7-AAD-GlyAB-CD45+ALDHbrCD34+ Cells
CB mononuclear cells (106 cells/mL) were stained with the ALDHbr Assay Kit and analyzed on a flow cytometer equipped with 488 nm and 625 nm lasers for detecting ALDEFLUOR™ (BAA), 7-AAD, APC, PE and PE-Cyanine5 fluorescence. The gating strategy for identifying (A) 7-AAD-GlyAB-CD45+ALDHbrCD34+ cells is shown using a fully stained sample (Sample) as described above. Staining of a (B) DEAB negative-control sample (Negative Control) is used to verify the placement of the ALDHbr gate. Gold lines depict the gates used at each stage to identify the desired cell types. Note: The gating of ALDHbr cells is greatly simplified if red blood cells (RBCs) are first removed from CB samples, prior to staining with ALDEFLUOR™. The ErythroClear™ Red Blood Cell Depletion Kit (Catalog #01739) is designed for the depletion of RBCs from up to 16 small volume CB samples.
Figure 3. The Frequencies of CD34+, ALDHbr and ALDHbrCD34+ Cells in CB Samples are Correlated with Colony Numbers as Measured in CFU Assays
CB mononuclear cells (n=23) were split into two parts. Half of each sample was stained with the ALDHbr Assay Kit and analyzed, as described in Figure 1. The percentages of (A) CD34+, (B) ALDHbr and (C) ALDHbrCD34+ cells in the 7-AAD-GlyAB-CD45+ population in each sample was determined. The other half of each sample was plated in a CFU assay using MethoCult™ Classic H4434 medium (Catalog #04434) and colonies were counted 14 days later. The frequency of CFUs was determined as a percentage of viable total nucleated cells (TNCs) for each sample. The table in (D) shows the number of cells from each subpopulation that contains 1 CFU. The data show that the frequency of ALDHbrCD34+ cells is highly correlated with the frequency of functional progenitor cells (R2: coefficient of determination).
Figure 4. The Percentage of Viable CD45+ Cells that Express CD34+ and/or ALDHbr Vary Among Different CB Samples
CB mononuclear cells (106 cells/mL) were stained and analyzed as described (Figure 1). Shown are the percentages of cells in the 7-AAD-GlyAB-CD45+ gated population that are CD34+, ALDHbr and ALDHbrCD34+. Each column represents the mean ± S.D. of three replicate assays for each CB sample (n=7).
Figure 5. The Analysis of the ALDHbrCD34+ Cell Subset is Consistent Across Different Flow Cytometers
A single CB mononuclear cell sample (106 cells/mL) was split into 3 parts, each of which was separately stained with the ALDHbr Assay Kit (see Figure 1 for protocol) and separately analyzed on three different flow cytometry instruments; a FACScan DXP6, FACSCalibur and Accuri C6. All instruments were equipped with 488 nm and 635 nm lasers for detecting ALDEFLUOR™ (BAA), 7-AAD, APC, PE and PE-Cyanine5 fluorescence. The frequency of ALDHbrCD34+ cells in each sample was determined as a percentage of the 7-AAD-GlyAB-CD45+ cell population measured by each instrument. There was no significant difference in the number of ALDHbrCD34+ cells measured by these flow cytometers (p > 0.05). Each column represents the mean ± S.D. for three replicates of the analyzed CB sample evaluated on each flow cytometer.
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。
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。
Thank you for your interest in IntestiCult™ Organoid Growth Medium (Human). Please provide us with your contact information and your local representative will contact you with a customized quote. Where appropriate, they can also assist you with a(n):
Estimated delivery time for your area
Product sample or exclusive offer
In-lab demonstration
种属 | Human |
---|
用于鉴定、评估和分离表达高水平ALDH的干细胞和祖细胞
用于ALDEFLUOR™试剂盒的附加分析缓冲液
用于分离单个核细胞的密度梯度离心液
扫描二维码或搜索微信号STEMCELLTech,即可关注我们的微信平台,第一时间接收丰富的技术资源和最新的活动信息。
如您有任何问题,欢迎发消息给STEMCELLTech微信公众平台,或与我们通过电话/邮件联系:400 885 9050 INFO.CN@STEMCELL.COM。