The BD Stemflow™ Human Neural Lineage Analysis Kit contains a combination of mouse monoclonal antibody conjugates for the analysis of neural differentiation cultures. The antibody specificities that are provided in this kit and the neural cell populations they can identify are listed in the table below. The antibody components of this kit can be used individually or in combinations of up to four. All Antibodies in this kit have been verified by western blot and image analysis. Additional antibody formats that can enable more complex panels are available at www.bdbiosciences.com. The kit also includes BD Cytofix™ Fixation Buffer and BD Phosflow™ Perm Buffer III.
Kit Components
Component
Description
Size
Vol. Per Test
Storage Buffer
51-9007227
PerCP-Cy™5.5 Mouse Anti-Sox2
25 Test
5 µl
Aqueous buffered solution containing
BSA and ≤ 0.09% sodium azide
51-9007228
Alexa Fluor® 647 Mouse Anti-GFAP
25 Test
5 µl
Aqueous buffered solution containing
BSA and ≤ 0.09% sodium azide
51-9007229
PE Mouse Anti-Doublecortin
25 Test
5 µl
Aqueous buffered solution containing
BSA and ≤ 0.09% sodium azide
51-9007230
Alexa Fluor® 647 Mouse Anti-Nestin
25 Test
5 µl
Aqueous buffered solution containing
BSA and ≤ 0.09% sodium azide
51-9007231
Alexa Fluor® 488 Mouse Anti-Human Ki-67
25 Test
5 µl
Aqueous buffered solution containing
BSA and ≤ 0.09% sodium azide
51-9007232
PerCP-Cy™5.5 Mouse Anti-Human Sox1
25 Test
5 µl
Aqueous buffered solution containing
BSA and ≤ 0.09% sodium azide
51-9007233
FITC Mouse Anti-Human CD44
25 Test
5 µl
Aqueous buffered solution containing
BSA and ≤ 0.09% sodium azide
51-9006607
Perm Buffer III
50 ml
51-9006276
Fixation Buffer
50 ml
Specificity
Clone
Molecule
Neural cell population identified
CD44
G44-26
H-CAM
Glial cells, astrocytes, astrocyte precursors
Ki-67
B56
Nuclear proliferation marker
All proliferating cell types
Doublecortin
30/Doublecortin
Neuroflilament
Immature post-mitotic neurons
Sox2
O30-678
Transcription factor
Glial cells, embryonic and neural stem cells
Sox1
N23-844
Transcription factor
Glial cells and neural stem cells
GFAP
1B4
Filament
Astrocytes
Nestin
25/Nestin
Filament
Glial cells, astrocytes, embryonic and neural stem cells
克隆号
(RUO)
应用
实验应用
Flow cytometry (Routinely Tested)
反应种属
Human (QC Testing)
目标/特异性
Neural Lineage Analysis Kit
文献
文献
研发参考(5)
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. (Methodology: Cell differentiation).
2. Itsykson P, Ilouz N, Turetsky T, Goldstein RS et al. Derivation of neural precursors from human embryonic stem cells in the presence of noggin. Mol Cell Neurosci. 2005; 30(1):24-36. (Methodology: Cell differentiation).
3. Koch P, Opitz T, Steinbeck JA, Ladewig J, Brüstle O. A rosette-type, self-renewing human ES cell-derived neural stem cell with potential for in vitro instruction and synaptic integration. Proc Natl Acad Sci U S A. 2009; 106(9):3225-3230. (Biology).
4. Reubinoff BE, Itsykson P, Turetsky T, et al. Neural progenitors from human embryonic stem cells. Nat Biotechnol. 2001; 19(12):1117-1118. (Methodology: Cell differentiation).
5. Yeo GW, Xu X, Liang TY, et al. Alternative splicing events identified in human embryonic stem cells and neural progenitors. PLoS Comput Biol. 2007; 3(10):1951-1967. (Biology).
参考图片
Neural Induction of H9 hESC: H9 Human embryonic stem cells (hESC) (Wicell Madison, WI) were cultured on mouse embryonic fibroblasts in hESC medium (DMEM:F12, 20% Knockout Serum Replacement (KOSR), non-essential amino acids, 20 mM glutamine) (Invitrogen), 1X penicillin/streptomycin (Lonza), bFGF (Cat. No. 354060). Cell cultures were treated with Dispase (Cat. No. 354235) and cell colonies were scraped and placed in hESC medium without FGF in low adhesion plates to form embroid bodies (EB). After 4 days the EBs were plated on BD™ Matrigel-coated plates (Cat. No. 354277) and placed in neural induction medium (DMEM:F12 (Invitrogen), 1X ITS supplement (Sigma), 250 ng/ml recombinant Noggin (R&D Systems) and penicillin/streptomycin (Lonza) and cultured for an additional 15 days. Differentiation of H9-derived NSC: H9 hESC-derived NSC were differentiated in neural differentiation medium (DMEM:F12, 0.5X B27, 0.5X N2 (Invitrogen), 1X penicillin/streptomycin (Lonza), 20 ng/ml BDNF, 20ng/ml GDNF (both from Peprotech) and 0.5 mM dibutyryl cyclic AMP (Sigma) for 28 days.
Differentiation of H9-derived NSC: H9 hESC-derived NSC were differentiated in neural differentiation medium (DMEM:F12, 0.5X B27, 0.5X N2 (Invitrogen), 1X penicillin/streptomycin (Lonza), 20 ng/ml BDNF, 20ng/ml GDNF (both from Peprotech) and 0.5 mM dibutyryl cyclic AMP (Sigma) for 11 days. Culture medium was replaced with AGM™ Astrocyte Growth Medium (Lonza) for an additional 16 days.