Traditional DNA/RNA isolation kits can be used to prepare genomic material for sequencing (RNA-seq, scRNA-seq, WGS, CHIP-seq etc). For some applications with large organoids, tissue specific DNA/RNA extraction kits are recommended. TRI Reagent (T9424) can be added to the dissociated organoids before RNA extraction and can be stored at -80 °C until future use.
SCC111
R-Spondin1 expressing 293T Cell Line
Human
Sinónimos:
RSPO1-expressing 293T cells
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$656.10
Precio de catálogo$729.00Ahorre 10 %Fecha estimada de envío12 de abril de 2025
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About This Item
$656.10
Precio de catálogo$729.00Ahorre 10 %Fecha estimada de envío12 de abril de 2025
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Nombre del producto
R-Spondin1 expressing 293T Cell Line, The R-Spondin1-Expressing 293T cell line produces RSPO1, a critical protein used to establish 3D organoid cultures from stomach, small intestine, colon, pancreas, and liver sources.
biological source
human
Quality Level
shipped in
ambient
General description
The 293T cell line is stably transfected to express mouse R-spondin1 protein tagged with N-terminus HA epitope tag and C-terminus mouse IgG2α Fc region. R-Spondin1 expressing 293T cell line can be used to produce either purified Rspo1 or Rspo1 conditioned media. The FC region and HA tags enable ease of purification and characterization. Cell line is not the published cell line from the Kuo lab, but rather a related cell line developed in the same lab using the reference protocol.
Cell Line Description
Application
Cell Culture
Quality
• Cells are tested negative for HPV-16, HPV-18, Hepatitis A, B, C, Herpesvirus type 6, 7, 8 and HIV-1 & 2 viruses by PCR.
• Cells are negative for mycoplasma contamination.
• Each lot of cells is genotyped by STR analysis to verify the unique identity of the cell line.
Storage and Stability
Disclaimer
Storage Class
10 - Combustible liquids
wgk_germany
WGK 1
flash_point_f
Not applicable
flash_point_c
Not applicable
Certificados de análisis (COA)
Busque Certificados de análisis (COA) introduciendo el número de lote del producto. Los números de lote se encuentran en la etiqueta del producto después de las palabras «Lot» o «Batch»
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How Do you Extract RNA/DNA from Organoids in Cultured in Matrigel?
1 answer-
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How are organoids cultured?
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Once isolated, organoids are cultured in specialized media embedded in a rich extracellular matrix (ECM) basement membrane extract hydrogel such as Growth Factor Reduced Matrigel. Media is replenished every other day and cells are passaged once per week (7-12 days) before cells become too large or necrotic. Cells can be passaged in either small clump fragments or as single cells using mechanical dissociation/enzyme-free passaging reagents. Depending on organoid type and confluency, split ratios of roughly 1:3-1:4 can be used when passaging organoids. For sensitive organoid types, ROCKi (Y27632) (SCM075) can be added during passaging to help in promoting cell viability.
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How do you stain organoids with antibodies?
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Organoids can be stained with antibodies for immunocytochemical (ICC)/Immunohistochemical (IHC) analysis using either whole-mount or paraffin embedding/sectioning techniques. It is critical to use antibodies prequalified for organoid analysis.
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What is the selection marker that is used in your RS Spondin expressing 293 cells. Is it neomycin resistance? What antibiotic should be used for selection pressure? Geneticin/G418?
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The cell line is selected by zeocin. It is not necessary to add zeocin to culture medium as the cell line is stable.
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