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SUFU KO cell line available to order. KO validated by Western blot. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, Homozygous: 11 bp deletion in exon 3.

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Images

Western blot - Human SUFU knockout HEK-293T cell line (AB267282), expandable thumbnail
  • Western blot - Human SUFU knockout HEK-293T cell line (AB267282), expandable thumbnail
  • Sanger Sequencing - Human SUFU knockout HEK-293T cell line (AB267282), expandable thumbnail

Key facts

Cell type
HEK-293T
Species or organism
Human
Tissue
Kidney
Form
Liquid
Knockout validation
Sanger Sequencing, Western blot
Mutation description
Knockout achieved by using CRISPR/Cas9, Homozygous: 11 bp deletion in exon 3

Alternative names

OTTHUMP00000020374, OTTHUMP00000020377, OTTHUMP00000020379, PRO1280, SU(F)U, SUFU negative regulator of hedgehog signaling, SUFUH, SUFUXL, SUFU_HUMAN, Su(fu), Suppressor of Fused, Suppressor of fused homolog, Suppressor of fused homolog (Drosophila)

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SUFU KO cell line available to order. KO validated by Western blot. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, Homozygous: 11 bp deletion in exon 3.

Key facts

Cell type
HEK-293T
Form
Liquid
Mutation description
Knockout achieved by using CRISPR/Cas9, Homozygous: 11 bp deletion in exon 3
Concentration
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Properties

Gene name
SUFU
Gene editing type
Knockout
Gene editing method
CRISPR technology
Knockout validation
Sanger Sequencing, Western blot
Zygosity
Homozygous

Quality control

STR analysis
CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX

Cell culture

Biosafety level
EU: 2 US: 2
Adherent/suspension
Adherent
Gender
Female

Handling procedures

Initial handling guidelines

Upon arrival, the vial should be stored in liquid nitrogen vapor phase and not at -80°C. Storage at -80°C may result in loss of viability.

1. Thaw the vial in 37°C water bath for approximately 1-2 minutes.
2. Transfer the cell suspension (0.8 mL) to a 15 mL/50 mL conical sterile polypropylene centrifuge tube containing 8.4 mL pre-warmed culture medium, wash vial with an additional 0.8 mL culture medium (total volume 10 mL) to collect remaining cells, and centrifuge at 201 x g (rcf) for 5 minutes at room temperature. 10 mL represents minimum recommended dilution. 20 mL represents maximum recommended dilution.
3. Resuspend the cell pellet in 5 mL pre-warmed culture medium and count using a haemocytometer or alternative cell counting method seed all remaining cells into a T25.
4. Incubate the culture at 37°C incubator with 5% CO2. Check the culture one day after revival and continue to check until 80% confluent. Media change can be given if needed.
5. Once confluent passage into an appropriate flask at a density of 2x104 cells/cm2. Seeding density is given as a guide only and should be scaled to align with individual lab schedules. Cultures should be monitored daily.

Subculture guidelines
  • All seeding densities should be based on cell counts gained by established methods.
  • A guide seeding density of 2x104 cells/cm2 is recommended.
  • Cells should be passaged when they have achieved 80-90% confluence.
Culture medium
DMEM (High Glucose) + 10% FBS
Cryopreservation medium
Cell Freezing Medium-DMSO Serum free media, contains 8.7% DMSO in MEM supplemented with methyl cellulose.

Storage

Shipped at conditions
Dry Ice
Appropriate short-term storage conditions
-196°C
Appropriate long-term storage conditions
-196°C

Notes

Recommended control: Human wild-type HEK293T cell line (ab255449). Please note a wild-type cell line is not automatically included with a knockout cell line order, if required please add recommended wild-type cell line at no additional cost using the code WILDTYPE-TMTK1.

We will provide viable cells that proliferate on revival.

This product is subject to limited use licenses from The Broad Institute and ERS Genomics Limited, and is developed with patented technology. For full details of the limited use licenses and relevant patents please refer to our limited use license and patent pages.

Supplementary info

This supplementary information is collated from multiple sources and compiled automatically.
Activity summary

SUFU also known as Suppressor of Fused is a regulatory protein with a mass of approximately 54 kDa. This protein serves as a significant component within the Hedgehog signaling pathway. SUFU is widely expressed in various tissues including brain heart and skeletal muscle. It acts as a negative regulator binding to and inhibiting the activity of other key proteins in the signaling cascade.

Biological function summary

SUFU modulates the activity of the transcription factors Gli1 Gli2 and Gli3 keeping them in the cytoplasm in their inactive form. It can form part of large protein complexes playing an important role in ensuring that signal transduction is tightly regulated. SUFU's function helps control the expression of target genes that are important for development and cellular differentiation. This regulation is essential during embryonic development and tissue maintenance in adults.

Pathways

SUFU has an important role in the Hedgehog signaling pathway and interacts with components like Smoothened (SMO) and Patched (PTCH). This pathway controls processes like cell growth and differentiation. By regulating the activity of Gli proteins SUFU influences how signals are transmitted within the pathway. SUFU can also interact with other proteins such as KIF7 further defining its place within the pathway's network.

Associated diseases and disorders

SUFU can contribute to the development of basal cell carcinoma and medulloblastoma when its function is disrupted. In cases where mutations affect SUFU regulation the Hedgehog pathway becomes aberrantly activated. This can lead to uncontrolled cell proliferation. SUFU's relationship with Gli proteins is significant in these conditions because of its role in modulating their activity directly impacting the onset and progression of these cancers.

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Product protocols

For this product, it's our understanding that no specific protocols are required. You can:

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