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BD Stemflow™ Human Neural Cell Sorting Kit

品牌： BD Pharmingen

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反应种属：

Human (Reactivity Confirmed in Development)

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简单描述

The BD Stemflow™ Neural Cell Isolation Kit was designed to allow the isolation of neural stem cells (NSCs) derived from human pluripotent stem cells or the isolation of neurons and glial cells from differentiated NSCs. Following a variety of established neural induction protocols, NSCs are sorted from the heterogeneous cell culture utilizing the included antibody conjugates to five cell surface markers. This method greatly reduces the amount of variability between NSC isolations as opposed to traditional methods like manual isolation. In addition, four of the five included conjugates can be used to isolate neurons from glia from differentiated NSCs. Kit Components Component Description Size Vol. Per Test Storage Buffer 51-9007757 PE Mouse Anti-Human CD24 20 Test 5 µl Aqueous buffered solution containing BSA and <0.09% sodium azide 51-9007758 PerCP-Cy™5.5 Mouse Anti-Human CD271 20 Test 5 µl Aqueous buffered solution containing BSA and <0.09% sodium azide 51-9007759 PerCP-Cy™5.5 Mouse Anti-Human CD44 20 Test 5 µl Aqueous buffered solution containing BSA and <0.09% sodium azide 51-9007760 PE-Cy™7 Mouse Anti-Human CD15 20 Test 5 µl Aqueous buffered solution containing BSA and <0.09% sodium azide 51-9007761 APC Mouse Anti-Human CD184 20 Test 5 µl Aqueous buffered solution containing BSA and <0.09% sodium azide 51-9007762 PE Mouse IgG1, κ Isotype Control 10 Test 5 µl Aqueous buffered solution containing BSA and <0.09% sodium azide 51-9007763 PerCP-Cy™5.5 Mouse IgG1, κ 10 Test 5 µl Aqueous buffered solution containing Isotype Control for 51-007758 BSA and <0.09% sodium azide 51-9007764 PerCP-Cy™5.5 Mouse IgG1, κ 10 Test 5 µl Aqueous buffered solution containing Isotype Control for 51-9007759 BSA and <0.09% sodium azide 51-9007765 PE-Cy™7 Mouse IgM, κ Isotype Control 10 Test 5 µl Aqueous buffered solution containing BSA and <0.09% sodium azide 51-9007766 APC Mouse IgG1, κ Isotype Control 10 Test 5 µl Aqueous buffered solution containing BSA and <0.09% sodium azide 51-9007767 Negative Control CompBead Plus 1.5 ml 1 drop Aqueous buffered solution containing BSA and <0.09% sodium azide 51-9007768 Anti-Mouse Ig, κ CompBead Plus 1.5 ml 1 drop Aqueous buffered solution containing BSA and <0.09% sodium azide Specificity Clone Molecule NSC Phenotype Neuron Phenotype Glial Phenotype CD15 HI98 X-hapten, SSEA-1 -/+ Low NA CD24 ML5 Heat Stable Antigen + + NA CD44 G44-26 H-CAM - - + CD184 12G5 CXCR4, Fusin + - + CD271 C40-1457 NGF-Receptor - NA NA

克隆号

(RUO)

应用

反应种属

Human (Reactivity Confirmed in Development)

目标/特异性

Neural Lineage Analysis Kit

文献

研发参考(2) 1. Chambers SM, Fasano CA, Papapetrou EP, Tomishima M, Sadelain M, Studer L. Highly efficient neural conversion of human ES and iPS cells by dual inhibition of SMAD signaling. Nat Biotechnol. 2009; 27(3):275-280. (Biology: Cell differentiation). 2. Yuan SH, Martin J, Elia J, et al. Cell-Surface Marker Signatures for the Isolation of Neural Stem Cells, Glia and Neurons Derived from Human Pluripotent Stem Cells. PLoS ONE. 6(3)(Methodology: Flow cytometry).

参考图片

Figure 1 Cell sorting of NSC from neural induction cultures: H9 hESC were differentiated into neural ectoderm using SFEB and dual SMAD inhibition. Please see Recommended Assay Procedure for more details. On day 20 of differentiation cells were processed as described above and sorted using the following gating strategy. Neural Stem Cell Sort (Gates based upon isotype controls): 1. Identify cells in FSC v SSC plot. (P1) a. Doublet discrimination should be applied after this step (P2) 2. Create a child gate from P2 that includes the CD184+ cells (P3). 3. Create a child gate from P3 that includes CD44- and CD271- cells (P4) 4. Create a child gate from P4 that in cludes the CD24+CD15+/- (NSC) cells Figure 2 Cell sorting of neurons and glia from differentiating NSC cultures: Previously sorted and expanded NSC derived from H9 hESC were differentiated for ~ 2.5 weeks in DMEM:F12+Glutamax, 1X B27, 1X N2 (Invitrogen), 1X P/S (Lonza), 20 ng/ml BDNF, 20ng/ml GDNF (both from Peprotech) and 0.5 mM dibutyryl cyclic AMP (Sigma). Cells were processed as described above and sorted using the following gating strategy. Neuron and Glia Sort (Gates based upon isotype controls): 1. Identify cells in FSC v SSC plot. (P1) a. Doublet discrimination should be applied after this step. (P2) b. Make sure to include the glia as they appear as a distinct scatter population in more mature cultures. 2. Create a child gates from P2 that includes CD44-CD184- cells (P3) or CD44+CD184+ cells (glia) 3. Create a child gate from P3 that includes the CD24+CD15- cells (Neurons)

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562271

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