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Human Three Germ Layer 3-Color Immunocytochemistry Kit

Catalog #: SC022
19 Citations Datasheet / COA / SDS
Contains six fluorochrome-conjugated anti-human antibodies: Ectoderm (SOX1, Otx-2); Mesoderm (Brachyury, HAND1); and Endoderm (GATA-4, SOX17)

Discontinued Product

SC022 has been discontinued.
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Verification of Germ Layer Markers in Differentiated Pluripotent Stem Cells.
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Citations (19)
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Human Three Germ Layer 3-Color Immunocytochemistry Kit Summary

Kit Summary

To analyze germ layer cells differentiated from human pluripotent stem cells.

Key Benefits

  • Verifies human stem cell pluripotency
  • Labels cells of the ectoderm, mesoderm, and endoderm
  • Can be used for simultaneous 3-color staining
 

 

Why Use Molecular Markers to Verify Human Stem Cell Pluripotency?

Research studies using embryonic stem (ES) or induced pluripotent stem (iPS) cells most often require weeks of cell culture before an experimental hypothesis can be tested.

Thus, the ability to verify pluripotency at an early stage in the experimental process is an important time saving step. Pluripotency status can be evaluated by the presence of stem cell markers and/or the ability of stem cells to differentiate into cells of the three germ layers. To determine if a cell is truly a pluripotent stem cell, it is important to verify its ability to differentiate into each of the three germ layers.

3-Color ICC using germ layer markers:

  • Evaluates the pluripotency status of the starting stem cell population.
  • Provides increased confidence in pluripotent status through the use of multiple markers for each germ layer.
  • Ensures efficient use of time and reagents by utilizing single-step staining.
  • Can be performed in randomly differentiated samples to simultaneously detect one marker from each germ layer.
 

 

Kit Contents

This kit contains the following fluorochrome-conjugated antibodies to analyze human pluripotent stem cells differentiated into cells of each germ layer:

Ectoderm

  • Anti-Human Otx2 NL557-Conjugated Goat IgG
  • Anti-Human SOX1 NL493-Conjugated Goat IgG

Mesoderm

  • Anti-Human Brachyury NL557-Conjugated Goat IgG
  • Anti-Human HAND1 NL637-Conjugated Goat IgG

Endoderm

  • Anti-Human GATA-4 NL493-Conjugated Goat IgG
  • Anti-Human SOX17 NL637-Conjugated Goat IgG

Each antibody is supplied as a 10X solution; enough for 25 assays when used in a 50 µL staining volume per assay.

Stability and Storage

Reagents are stable for 12 months from date of receipt when stored in the dark at 2° C to 8°.

Specifications

Source
N/A
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

Product Datasheets

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Scientific Data

Cell Differentiation/ Maturation Verification of Germ Layer Markers in Differentiated Pluripotent Stem Cells. View Larger

Verification of Germ Layer Markers in Differentiated Pluripotent Stem Cells. iPS2 human induced pluripotent stem cells were differentiated into each of the three germ layers using the base media and differentiation supplements provided in the Human Pluripotent Stem Cell Functional Identification Kit (Catalog # SC027). Germ layer differentiation was subsequently verified using the six fluorochrome-conjugated antibodies provided in the Human Three Germ Layer 3-Color Immunocytochemistry Kit (Catalog # SC022). Ectoderm differentiated cells were simultaneously stained with NorthernLights (NL) fluorochrome NL557-conjugated Otx2 (red) and NL493-conjugated SOX1 (green). Mesoderm differentiated cells were simultaneously stained with NL557-conjugated Brachyury (red) and NL637-conjugated HAND1 (green). Endoderm differentiated cells were simultaneously stained with NL637-conjugated SOX17 (red) and NL493-conjugated GATA-4 (green). All nuclei were counterstained with DAPI (blue). Nuclear expression of each marker was detected in their respective cell lineage.

Cell Differentiation/ Maturation Random Differentiation within a BG01V Human Embryonic Stem Cell-derived Embryoid Body. View Larger

Random Differentiation within a BG01V Human Embryonic Stem Cell-derived Embryoid Body. BG01V human embryonic stem cell-derived embryoid bodies were grown on plates coated with Cultrex®Stem Cell Qualified Basement Membrane Extract (Catalog # 3434-005-02). Random differentiation was detected by simultaneous 3-color staining using three fluorochrome-conjugated antibodies provided in the Human Three Germ Layer 3-Color Immunocytochemistry Kit (Catalog # SC022). Ectoderm derivatives were detected with NorthernLights (NL) fluorochrome NL557-conjugated Otx2 (red), mesoderm differentiated cells with NL637-conjugated HAND1 (white), and ectoderm derivatives with NL493-conjugated GATA-4 (green). The nuclei were counterstained with DAPI (blue).

Citations for Human Three Germ Layer 3-Color Immunocytochemistry 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.

19 Citations: Showing 1 - 10
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  1. Generation of iPSC lines from three Laing distal myopathy patients with a recurrent MYH7 p.Lys1617del variant
    Authors: Clayton, JS;Vo, C;Crane, J;Scriba, CK;Saker, S;Larmonier, T;Malfatti, E;Romero, NB;Ravenscroft, G;Laing, NG;Taylor, RL;
    Stem cell research  2024-07-09
  2. Comparison of osteoclast differentiation protocols from human induced pluripotent stem cells of different tissue origins
    Authors: Blümke, A;Ijeoma, E;Simon, J;Wellington, R;Purwaningrum, M;Doulatov, S;Leber, E;Scatena, M;Giachelli, CM;
    Research square  2023-07-07
  3. Generation of two induced pluripotent stem cell lines (CHOCi002-A and CHOCi003-A) from Pompe disease patients with compound heterozygous mutations in the GAA gene
    Authors: Christensen, C;Heckman, P;Rha, A;Kan, SH;Harb, J;Wang, R;
    Stem cell research  2023-05-06
  4. Generation of the human erythroblast-derived iPSC line UBTi001-A
    Authors: A Avdili, L Rohrhofer, M Auer, C Bernecker, P Schlenke, I Dorn
    Stem Cell Research, 2022-09-07;64(0):102910.  2022-09-07
  5. Generation of two isogenic induced pluripotent stem cell lines from a 1-month-old nemaline myopathy patient harbouring a homozygous recessive c.121C�>�T (p.Arg39Ter) variant in the ACTA1 gene
    Authors: IS Suleski, R Smith, C Vo, CK Scriba, S Saker, T Larmonier, E Malfatti, NB Romero, PJ Houweling, KJ Nowak, NG Laing, RL Taylor, JS Clayton
    Stem Cell Research, 2022-06-06;63(0):102830.  2022-06-06
  6. Generation of an induced pluripotent stem cell line from a 3-month-old nemaline myopathy patient with a heterozygous dominant c.515C�>�A (p.Ala172Glu) variant in the ACTA1 gene
    Authors: JS Clayton, I Suleski, C Vo, R Smith, CK Scriba, S Saker, T Larmonier, E Malfatti, NB Romero, PJ Houweling, KJ Nowak, G Ravenscrof, NG Laing, RL Taylor
    Oncogene, 2022-06-06;63(0):102829.  2022-06-06
  7. Generation and characterization of an endogenously tagged SPG11-human iPSC line by CRISPR/Cas9 mediated knock-in
    Authors: L Krumm, T Pozner, J Kaindl, M Regensburg, C Günther, S Turan, R Asadollahi, A Rauch, B Winner
    Stem Cell Research, 2021-08-26;56(0):102520.  2021-08-26
  8. Generation of disease-specific and CRISPR/Cas9-mediated gene-corrected iPS cells from a patient with adult progeria Werner syndrome
    Authors: H Kato, Y Maezawa, Y Ouchi, N Takayama, M Sone, K Sone, A Takada-Wat, K Tsujimura, M Koshizaka, S Nagasawa, H Saitoh, M Ohtaka, M Nakanishi, H Tahara, A Shimamoto, A Iwama, K Eto, K Yokote
    Stem Cell Research, 2021-04-23;53(0):102360.  2021-04-23
  9. Generation of two isogenic induced pluripotent stem cell lines from a 4-month-old severe nemaline myopathy patient with a heterozygous dominant c.553C�>�A (p.Arg183Ser) variant in the ACTA1 gene
    Authors: JS Clayton, CK Scriba, NB Romero, E Malfatti, S Saker, T Larmonier, KJ Nowak, G Ravenscrof, NG Laing, RL Taylor
    Stem Cell Research, 2021-02-26;53(0):102273.  2021-02-26
  10. Generation of human induced pluripotent stem cells (hIPSCs) from sialidosis types I and II patients with pathogenic neuraminidase 1 mutations
    Authors: MJ Han, I Annunziata, J Weesner, Y Campos, M Salie, C O'Reilly, A d'Azzo
    Stem Cell Res, 2020-05-06;46(0):101836.  2020-05-06
  11. Generation of Best disease-derived induced pluripotent stem cell line (FRIMOi006-A) carrying a novel dominant mutation in BEST1 gene
    Authors: J Domingo-Pr, M Riera, V Abad-Moral, S Ruiz-Nogal, B Corcostegu, E Pomares
    Stem Cell Res, 2019-09-04;40(0):101570.  2019-09-04
  12. Generation of an induced pluripotent stem cell line (FRIMOi007-A) derived from an incomplete achromatopsia patient carrying a novel homozygous mutation in PDE6C gene
    Authors: J Domingo-Pr, V Abad-Moral, M Riera, R Navarro, B Corcostegu, E Pomares
    Stem Cell Res, 2019-09-04;40(0):101569.  2019-09-04
  13. Establishment of an induced pluripotent stem cell line (FRIMOi005-A) derived from a retinitis pigmentosa patient carrying a dominant mutation in RHO gene
    Authors: J Domingo-Pr, M Riera, A Burés-Jels, B Corcostegu, E Pomares
    Stem Cell Res, 2019-05-22;38(0):101468.  2019-05-22
  14. Generation of two iPS cell lines (FRIMOi003-A and FRIMOi004-A) derived from Stargardt patients carrying ABCA4 compound heterozygous mutations
    Authors: M Riera, A Patel, A Burés-Jels, B Corcostegu, S Chang, E Pomares, B Corneo, JR Sparrow
    Stem Cell Res, 2019-02-13;36(0):101389.  2019-02-13
  15. Generation of an induced pluripotent stem cell line (FRIMOi002-A) from a retinitis pigmentosa patient carrying compound heterozygous mutations in USH2A gene
    Authors: M Riera, A Patel, B Corcostegu, S Chang, B Corneo, JR Sparrow, E Pomares
    Stem Cell Res, 2019-01-17;35(0):101386.  2019-01-17
  16. Establishment and characterization of an iPSC line (FRIMOi001-A) derived from a retinitis pigmentosa patient carrying PDE6A mutations
    Authors: M Riera, A Patel, B Corcostegu, S Chang, JR Sparrow, E Pomares, B Corneo
    Stem Cell Res, 2019-01-17;35(0):101385.  2019-01-17
  17. Disruption of mesoderm formation during cardiac differentiation due to developmental exposure to 13-cis-retinoic acid
    Authors: Q Liu, K Van Bortle, Y Zhang, MT Zhao, JZ Zhang, BS Geller, JJ Gruber, C Jiang, JC Wu, MP Snyder
    Sci Rep, 2018-08-28;8(1):12960.  2018-08-28
  18. Susceptibility of Human Oral Squamous Cell Carcinoma (OSCC) H103 and H376 cell lines to Retroviral OSKM mediated reprogramming
    Authors: ND Verusingam, SK Yeap, H Ky, IC Paterson, SP Khoo, SK Cheong, AHK Ong, T Kamarul
    PeerJ, 2017-04-13;5(0):e3174.  2017-04-13
  19. Generation of human iPSCs from urine derived cells of a patient with a novel homozygous PAI-1 mutation
    Stem Cell Res, 2016-11-09;17(3):657-660.  2016-11-09

FAQs

  1. In Figure 1 on the Datasheet for Catalog # SC022, why is the staining produced by the antibody conjugated to NL637 dye green in one image and red in another image?

    • NL637 is assigned a different color (green) than its natural color in one of the the images in order to obtain good contrast with the co-staining done for another marker in the same sample. Red is the natural color for NL637.

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