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H/M Pluripotent Stem Cell Multi-Color Flow Cytometry Kit

Catalog #: FMC001
13 Citations Datasheet / COA / SDS
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FMC001
Verification of Human BG01V Embryonic Stem Cell Pluripotency by Multi-Color Flow Cytometry.
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H/M Pluripotent Stem Cell Multi-Color Flow Cytometry Kit Summary

Kit Summary

For the verification of stem cell pluripotency using four established markers.

Why is it Important to Verify Embryonic Stem Cell Pluripotency Using Multiple Established Markers?

Identifying the sources of experimental variability is an important consideration in stem cell research where experiments are costly and time-consuming. A potential source of significant variability arises from the starting population of stem cells which can undergo phenotypic changes in culture.   See Details

Changes in stem cell potency over time can give rise to large inter-assay errors and/or contradictory data. The Human/Mouse Pluripotent Stem Cell Multi-Color Flow Cytometry Kit offers users an efficient and quantitative method to verify the pluripotency of cells by flow cytometry. Data obtained using this kit can identify and minimize experimental errors introduced by variations in the starting population of cells.

The Human/Mouse Pluripotent Stem Cell Multi-Color Flow Cytometry Kit:

  • Efficiently verifies stem cell pluripotency using four markers of pluripotency.
  • Defines the starting population to reduce experimental variability.
  • Does not require the use of secondary antibodies.
  • Includes enough reagents to perform 25 assays.

Kit Components

The Human/Mouse Pluripotent Stem Cell Multi-Color Flow Cytometry Kit includes four fluorochrome-conjugated primary antibodies, isotype controls and buffers to fix, permeabilize, and wash cells.   See Details

  • SOX2-PE (Clone 245610; mouse IgG2A)
  • Oct-3/4-APC (Clone 240408; rat IgG2B)
  • SSEA-1-PerCP (Clone MC-480; mouse IgM)
  • SSEA-4-CFS (Clone MC-813-70; mouse IgG3)
  • Mouse IgM Isotype Control
  • Mouse IgG3 Isotype Control
  • Rat IgG2B Isotype Control
  • Mouse IgG2A Isotype Control
  • Fixation/Permeabilization Buffer (with 1% formaldehyde, 8.9% saponin, and <0.05% sodium azide)
  • Permeabilization/Wash Buffer (with 10% saponin and 0.05% sodium azide)
  • Includes enough reagents to perform 25 assays

Stability and Storage

Store at 2 °C to 8 °C in the dark. Use within 6 months of receipt.

Specifications

Shipping Conditions
The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.
Storage
Store the unopened product at 2 - 8 °C. Do not use past expiration date.
Species
Human Mouse

Product Datasheets

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Scientific Data

N/A Verification of Human BG01V Embryonic Stem Cell Pluripotency by Multi-Color Flow Cytometry. View Larger

Verification of Human BG01V Embryonic Stem Cell Pluripotency by Multi-Color Flow Cytometry. BG01V human embryonic stem cells were stained using reagents included in the Human/Mouse Pluripotent Stem Cell Multi-Color Flow Cytometry Kit (Catalog # FMC001). Cells were simultaneously analyzed for expression of pluripotent markers including SSEA-1, SSEA-4, Oct-3/4, and SOX2 by flow cytometry. A. Flow cytometry data shows that 91.9% of BG01V human embryonic stem cells are positive for both Oct-3/4 and SSEA4 expression. B. Flow cytometry data shows that 88.5% of BG01V human embryonic stem cells are positive for SSEA-4 and negative for SSEA-1, a phenotype consistent with human embryonic stem cells. C. Flow cytometric analysis shows that BG01V human embryonic stem cells express the pluripotent marker SOX2.

N/A Verification of Mouse D3 Embryonic Stem Cell Pluripotency by Multi-Color Flow Cytometry. View Larger

Verification of Mouse D3 Embryonic Stem Cell Pluripotency by Multi-Color Flow Cytometry. Mouse D3 embryonic stem cells were stained using reagents included in the Human/Mouse Embryonic Stem Cell Multi-Color Flow Cytometry Kit (Catalog # FMC001). Cells were analyzed for expression of pluripotent markers including SSEA-1, SSEA-4, Oct-3/4, and SOX2 by flow cytometry. A. Flow cytometric analysis shows that 91.1% of mouse D3 embryonic stem cells are positive for both Oct-3/4 and SSEA1 expression. B. Flow cytometric analysis data shows that 82.6% of mouse D3 embryonic stem cells are positive for SSEA-1 and negative for SSEA-4 a phenotype consistent with mouse embryonic stem cells. C. Flow cytometric analysis shows that mouse D3 embryonic stem cells express the pluripotent marker SOX2.

&#9888; WARNING: This product can expose you to chemicals including formaldehyde and methanol, which are known to the State of California to cause cancer and reproductive toxicity with developmental effects. For more information, go to www.P65Warnings.ca.gov.

Assay Procedure

 

>Refer to the product datasheet for complete product details.

Briefly, stem cell pluripotency is assessed by flow cytometry using the following procedure:

  • Fix and permeabilize cells using Fixation/Permeabilization Buffer
  • Stain cells with fluorochrome-conjugated antibodies or isotype controls
  • Wash and resuspend cells in PBS
  • Analyze samples by flow cytometry
ARY020 Protocol Graphic Summary
View Graphic Protocol
 

Reagents Provided

Reagents Supplied in the Human/Mouse Embryonic Stem Cell Multi-Color Flow Cytometry Kit (Catalog # FMC001)

  • SOX2-PE (Clone 245610; mouse IgG2A)
  • Oct-3/4-APC (Clone 240408; rat IgG2B)
  • SSEA-1-PerCP (Clone MC-480; mouse IgM)
  • SSEA-4-CFS (Clone MC-813-70; mouse IgG3)
  • Mouse IgM Isotype Control
  • Mouse IgG3 Isotype Control
  • Rat IgG2B Isotype Control
  • Mouse IgG2A Isotype Control
  • Fixation/Permeabilization Buffer (with 1% formaldehyde, 8.9% saponin, and <0.05% sodium azide)
  • Permeabilization/Wash Buffer (with 10% saponin and 0.05% sodium azide)
  • Includes enough reagents to perform 25 assays

Other Supplies Required

  • PBS or Hanks’ Balanced Salt Solution (HBSS)

Procedure Overview

Intracellular Staining Protocol with Simultaneous Fixation/Permeabilization

  1. Harvest cells and wash 2 times with PBS or HBSS.
wash with PBS or HBSS
  1. Resuspend cells in Fixation/Permeabilization Buffer (approximately 5 x 105 cells/0.5 mL) and transfer to 5 mL flow cytometry tubes.
  2. Incubate cells at room temperature for 30 minutes, vortexing intermittently.
resuspend cells in buffer and transfer to flow cytometry tubes
  1. Centrifuge samples at 300 x g for 5 minutes.
  2. Resuspend the pellet in 100-200 μL of Permeabilization/Wash Buffer.
resuspend the pellet in wash buffer
  1. Add 10 µL of each fluorochrome-conjugated antibody or the corresponding isotype control antibody.
  2. Incubate the samples at room temperature for 30-40 minutes in the dark.
add fluorchorome-conjugated antibody or isotype control antibody
  1. Wash the cells in Permeabilization/Wash Buffer.
  2. Resuspend the cells in 200-400 μL of PBS.
wash cells in buffer and resuspend cells in PBS
  1. Analyze the expression of pluripotent markers simultaneously by flow cytometry.
Analyze the expression of pluripotent markers simultaneously by flow cytometry

Citations for H/M Pluripotent Stem Cell Multi-Color Flow Cytometry Kit

R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

13 Citations: Showing 1 - 10
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  1. Generation of the human pluripotent stem cell lines KUMi005-A from a patients with multiple myeloma
    Authors: JH Kim, JH Hong, KW Kang, BH Lee, Y Park, BS Kim
    Stem Cell Research, 2022-10-13;65(0):102939.  2022-10-13
  2. Generation of human induced iPSC KUMi003-A from acute promyelocytic leukemia (APL) M3
    Authors: JH Kim, KW Kang, BH Lee, Y Park, BS Kim
    Oncogene, 2022-07-09;63(0):102861.  2022-07-09
  3. Generation of a human induced pluripotent stem cell line KUMi006 from a patient with multiple myeloma
    Authors: JH Hong, JH Kim, KW Kang, BH Lee, Y Park, BS Kim
    Stem Cell Research, 2022-03-28;61(0):102767.  2022-03-28
  4. Generation and characterization of a human iPSC line SANi006-A from a Gray Platelet Syndrome patient
    Authors: CEM Aarts, E Varga, S Webbers, J Geissler, M von Linder, TW Kuijpers, E van den Ak
    Stem Cell Research, 2021-06-29;55(0):102443.  2021-06-29
  5. Generation and characterization of a human iPSC line SANi007-A from a patient with a heterozygous dominant mutation in ELANE
    Authors: CEM Aarts, E Varga, S Webbers, J Geissler, M von Linder, TW Kuijpers, E van den Ak
    Stem Cell Research, 2021-06-27;55(0):102440.  2021-06-27
  6. Generation and characterization of a human iPSC line SANi008-A from a Ch�diak-Higashi Syndrome patient
    Authors: CEM Aarts, E Varga, S Webbers, J Geissler, M von Linder, TW Kuijpers, E van den Ak
    Stem Cell Research, 2021-06-24;55(0):102442.  2021-06-24
  7. Generation and characterization of a control and patient-derived human iPSC line containing the Hermansky Pudlak type 2 (HPS2) associated heterozygous compound mutation in AP3B1
    Authors: CEM Aarts, E Karampini, T Wüst, S Webbers, E Varga, J Geissler, J Voorberg, M von Linder, R Bierings, E van den Ak, TW Kuijpers
    Stem Cell Research, 2021-06-23;54(0):102444.  2021-06-23
  8. Modelling heme-mediated brain injury associated with cerebral malaria in human brain cortical organoids
    Authors: A Harbuzariu, S Pitts, JC Cespedes, KO Harp, A Nti, AP Shaw, M Liu, JK Stiles
    Sci Rep, 2019-12-16;9(1):19162.  2019-12-16
  9. Generation of the induced human pluripotent stem cell lines CSSi009-A from a patient with a GNB5 pathogenic variant, and CSSi010-A from a CRISPR/Cas9 engineered GNB5 knock-out human cell line
    Authors: N Malerba, P Benzoni, GM Squeo, R Milanesi, F Giannetti, LG Sadleir, G Poke, B Augello, AI Croce, A Barbuti, G Merla
    Stem Cell Res, 2019-08-22;40(0):101547.  2019-08-22
  10. Efficient Generation of Non-Integration and Feeder-Free Induced Pluripotent Stem Cells from Human Peripheral Blood Cells by Sendai Virus
    Authors: H Ye, Q Wang
    Cell. Physiol. Biochem., 2018-10-24;50(4):1318-1331.  2018-10-24
  11. Derivation of human induced pluripotent stem cell line EURACi004-A from skin fibroblasts of a patient with Arrhythmogenic Cardiomyopathy carrying the heterozygous PKP2 mutation c.2569_3018del50
    Authors: B Ermon, CB Volpato, G Cattelan, R Silipigni, M Di Segni, C Cantaloni, M Casella, PP Pramstalle, G Pompilio, E Sommariva, V Meraviglia, A Rossini
    Stem Cell Res, 2018-09-06;32(0):78-82.  2018-09-06
  12. Microfabric Vessels for Embryoid Body Formation and Rapid Differentiation of Pluripotent Stem Cells
    Sci Rep, 2016-08-10;6(0):31063.  2016-08-10
  13. Differentiation and transplantation of functional pancreatic beta cells generated from induced pluripotent stem cells derived from a type 1 diabetes mouse model.
    Authors: Jeon, Kilsoo, Lim, Hyejin, Kim, Jung-Hyu, Thuan, Nguyen V, Park, Seung Hw, Lim, Yu-Mi, Choi, Hye-Yeon, Lee, Eung-Ryo, Kim, Jin-Hoi, Lee, Myung-Sh, Cho, Ssang-Go
    Stem Cells Dev, 2012-06-01;21(14):2642-55.  2012-06-01

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