Human SUFU knockout HEK-293T cell line
- Advanced Validation
- What is this?
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SUFU KO cell line available to order. KO validated by Western blot. Free of charge wild type control available. Knockout achieved by using CRISPR/Cas9, Homozygous: 11 bp deletion in exon 3. To order both knockout and wild-type control cells: select '2 x 1000000 Cells/vial'. To order only knockout cells: select '1000000 Cells/vial'.
View 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)
- WB
Lab
Western blot - Human SUFU knockout HEK-293T cell line (AB267282)
Blocking and diluting buffer and concentration : Intercept® (TBS) Blocking Buffer diluted with an equal volume of 0.1% TBS.
Lanes 1-3 : Merged signal (red and green). Green - ab259975 observed at 53 kDa. Red - loading control ab8245 (Mouse monoclonal [6C5] to GAPDH) observed at 36 kDa.
ab253183 Anti-SUFU antibody [EPR23821-101] was shown to specifically react with SUFU in wild-type HEK293T cells. Loss of signal was observed when knockout cell line ab267282 (knockout cell lysate ab257718) was used. Wild-type and SUFU knockout samples were subjected to SDS-PAGE.
ab253183 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated at 4° overnight at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 10000 dilution for 1 hour at room temperature before imaging.
All lanes:
Western blot - Anti-SUFU antibody [EPR23821-101] (<a href='/en-us/products/primary-antibodies/sufu-antibody-epr23821-101-ab259975'>ab259975</a>) at 1/1000 dilution
Lane 1:
Wild-type HEK293T (human embryonic kidney epithelial cell), whole cell lysate at 20 µg
Lane 2:
SUFU knockout HEK293T (human embryonic kidney epithelial cell), whole cell lysate at 20 µg
Lane 2:
Western blot - Human SUFU knockout HEK-293T cell line (ab267282)
Lane 3:
LNCaP (human prostate carcinoma epithelial cell), whole cell lysate at 20 µg
Secondary
All lanes:
Goat Anti-Rabbit IgG H&L (IRDye® 800CW) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-irdye-800cw-preadsorbed-ab216773'>ab216773</a>) and Goat Anti-Mouse IgG H&L (IRDye® 680RD) (<a href='/en-us/products/secondary-antibodies/goat-mouse-igg-h-l-irdye-680rd-preadsorbed-ab216776'>ab216776</a>) at 1/10000 dilution
Predicted band size: 53 kDa
Observed band size: 53 kDa
false
- WB
Lab
Western blot - Human SUFU knockout HEK-293T cell line (AB267282)
Lanes 1-3 : Merged signal (red and green). Green - ab28083 observed at 58 kDa. Red - loading control ab8245 observed at 36 kDa.
ab28083 Anti-SUFU antibody was shown to specifically react with SUFU in wild-type HEK293T cells. Loss of signal was observed when knockout cell line ab267282 (knockout cell lysate ab257718) was used. Wild-type and SUFU knockout samples were subjected to SDS-PAGE. ab28083 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated at room temperature for 2.5 hours at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
All lanes:
Western blot - Anti-SUFU antibody (<a href='/en-us/products/primary-antibodies/sufu-antibody-ab28083'>ab28083</a>) at 1/1000 dilution
Lane 1:
Wild-type HEK293T cell lysate at 20 µg
Lane 2:
SUFU knockout HEK293T cell lysate at 20 µg
Lane 2:
Western blot - Human SUFU knockout HEK-293T cell line (ab267282)
Lane 3:
LNCaP cell lysate at 20 µg
Secondary
All lanes:
Western blot - Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-irdye-800cw-preadsorbed-ab216773'>ab216773</a>) at 1/10000 dilution
Predicted band size: 53 kDa
Observed band size: 58 kDa
false
- Sanger seq
Unknown
Sanger Sequencing - Human SUFU knockout HEK-293T cell line (AB267282)
Homozygous : 11 bp deletion in exon3
- ICC/IF
Lab
Immunocytochemistry/ Immunofluorescence - Human SUFU knockout HEK-293T cell line (AB267282)
Immunofluorescent analysis of 100% methanol-fixed, 0.1% TritonX-100 permeabilized Wildtype HEK-293T (human embryonic kidney epithelial cell), ab255449 and SUFU KO HEK-293T (SUFU knockout HEK-293T), ab267282 cells labelling SUFU with ab325424 at 1/200 (2.445 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 dilution (Green).
Confocal image showing mainly cytoplasmic staining in wildtype HEK-293T cells (shown in green), showing no staining in SUFU knockout HEK-293T cells. The counterstain was observed in magenta. Nuclear DNA was labelled with DAPI (shown in blue). Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).
ab7291 Anti-alpha Tubulin mouse monoclonal antibody was used to counterstain tubulin at 1/1000 dilution, followed by ab150120 Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) at 1/1000 dilution (Magenta).
Reactivity data
Product details
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.
What's included?
Properties and storage information
Gene name
Gene editing type
Gene editing method
Knockout validation
Zygosity
Shipped at conditions
Appropriate short-term storage conditions
Appropriate long-term storage conditions
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.
Supplementary information
This supplementary information is collated from multiple sources and compiled automatically.
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.
Quality control
STR analysis
CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
Biosafety level
EU: 2 US: 2
Adherent/suspension
Adherent
Gender
Female
Complementary Products
Primary Antibodies
AB259975
Anti-SUFU antibody [EPR23821-101]
RabMAb|Recombinant|KO Validated|Trial Size
![Flow Cytometry (Intracellular) - Anti-SUFU antibody [EPR23821-101] (AB259975)](https://content.abcam.com/products/images/sufu-antibody-epr23821-101-ab259975--flow-cytometry-intracellular-img13459.jpg)
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Product promise
Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.
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