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ALDHbr测定试剂盒

用于人脐带血CD34+和ALDHbr细胞的检测
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产品号 #（选择产品）

产品号 #01711_C

用于人脐带血CD34+和ALDHbr细胞的检测

产品优势

  • 在< 1天内定量原始hsc
  • 优化检测CB中ALDHbrCD34+细胞
  • 枚举CD34+细胞群中存活的、代谢活跃的原始细胞

产品组分包括

  • 1 x ALDEFLUOR™套件（目录#01700）
    • ALDEFLUOR™试剂
    • ALDEFLUOR™DEAB试剂
    • ALDEFLUOR™测定缓冲液
    • 盐酸
    • DMSO溶液
  • 2 × CD34+细胞检测试剂盒
    • 7-AAD活性染料
    • 抗人CD45抗体，克隆HI30， PE
    • 抗人CD34抗体，克隆581
    • 抗人CD235ab（糖蛋白AB）抗体，克隆HIR2， PE-Cyanine5
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概述

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
 

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Data Figures

General ALDHbr Assay Procedure

Figure 1. General ALDHbr Assay Procedure

For a detailed protocol please refer to the ALDHbr Assay Kit Product Information Sheet (PIS)

Gating Strategy to Identify 7-AAD-GlyAB-CD45+ALDHbrCD34+ Cells

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.

The Frequencies of CD34+, ALDHbr and ALDHbrCD34+ Cells in CB Samples are Correlated with Colony Numbers as Measured in CFU Assays

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).

The Percentage of Viable CD45+ Cells that Express CD34+ and/or ALDHbr Vary Among Different CB Samples

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).

The Analysis of the ALDHbrCD34+ Cell Subset is Consistent Across Different Flow Cytometers

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.

Protocols and Documentation

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

Document Type
Product Name
Catalog #
Lot #
Language
Document Type
Product Information Sheet 1
Product Name
ALDHbr Assay Kit
Catalog #
01711
Lot #
All
Language
English
Document Type
Product Information Sheet 2
Product Name
ALDHbr Assay Kit
Catalog #
01711
Lot #
All
Language
English
Document Type
Safety Data Sheet 1
Product Name
ALDHbr Assay Kit
Catalog #
01711
Lot #
All
Language
English
Document Type
Safety Data Sheet 2
Product Name
ALDHbr Assay Kit
Catalog #
01711
Lot #
All
Language
English
Document Type
Safety Data Sheet 3
Product Name
ALDHbr Assay Kit
Catalog #
01711
Lot #
All
Language
English
Document Type
Safety Data Sheet 4
Product Name
ALDHbr Assay Kit
Catalog #
01711
Lot #
All
Language
English
Document Type
Safety Data Sheet 5
Product Name
ALDHbr Assay Kit
Catalog #
01711
Lot #
All
Language
English
Document Type
Safety Data Sheet 6
Product Name
ALDHbr Assay Kit
Catalog #
01711
Lot #
All
Language
English
Document Type
Safety Data Sheet 7
Product Name
ALDHbr Assay Kit
Catalog #
01711
Lot #
All
Language
English
Document Type
Safety Data Sheet 8
Product Name
ALDHbr Assay Kit
Catalog #
01711
Lot #
All
Language
English
Document Type
Safety Data Sheet 9
Product Name
ALDHbr Assay Kit
Catalog #
01711
Lot #
All
Language
English

Applications

This product is designed for use in the following research area(s) as part of the highlighted workflow stage(s). Explore these workflows to learn more about the other products we offer to support each research area.

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Species Human
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