Human AES (Amino-terminal enhancer of split) knockout HCT116 cell line
- Advanced Validation
- What is this?
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TLE5 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: 1 bp deletion in exon 2.
View Alternative Names
AES_HUMAN, Aes 1, Aes 2, Amino enhancer of split, Amino-terminal enhancer of split, ESP-1, GRG, GRG protein, Gp130-associated protein GAM, Grg-5, Groucho-related protein 5, Protein ESP 1, Protein GRG, aes
- WB
Lab
Western blot - Human AES (Amino-terminal enhancer of split) knockout HCT116 cell line (AB266886)
Lanes 1-4 : Merged signal (red and green). Green - ab137060 observed at 21 kDa. Red - loading control ab8245 observed at 36 kDa.
ab137060 Anti-Amino-terminal enhancer of split/AES antibody [EPR8385] was shown to specifically react with Amino-terminal enhancer of split/AES in wild-type HCT cells. Loss of signal was observed when knockout cell line ab266886 (knockout cell lysate ab257818) was used. Wild-type and Amino-terminal enhancer of split/AES knockout samples were subjected to SDS-PAGE. ab137060 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C 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-Amino-terminal enhancer of split/AES antibody [EPR8385] (<a href='/en-us/products/primary-antibodies/amino-terminal-enhancer-of-split-aes-antibody-epr8385-ab137060'>ab137060</a>) at 1/1000 dilution
Lane 1:
Wild-type HCT 116 (Human colorectal carcinoma cell line) whole cell lysate at 20 µg
Lane 2:
AES knockout HCT 116 (Human colorectal carcinoma cell line) whole cell lysate at 20 µg
Lane 2:
Western blot - Human AES (Amino-terminal enhancer of split) knockout HCT116 cell line (ab266886)
Lane 3:
HEK-293T (Human epithelial cell line from embryonic kidney transformed with large T antigen) whole cell lysate at 20 µg
Lane 4:
PC3 (Human prostate adenocarcinoma cell line) whole 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: 21 kDa
Observed band size: 21 kDa
false
- Sanger seq
Unknown
Sanger Sequencing - Human AES (Amino-terminal enhancer of split) knockout HCT116 cell line (AB266886)
Homozygous : 1 bp deletion in exon2
- Cell Culture
Lab
Cell Culture - Human AES (Amino-terminal enhancer of split) knockout HCT116 cell line (AB266886)
Representative images AES knockout HCT116 cells, low and high confluency examples (top left and right respectively) and wild-type HCT116 cells, low and high confluency (bottom left and right respectively) showing typical adherent, epithelial-like morphology. Images were captured at 10X magnification using a EVOS M5000 microscope.
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
McCoY5a + 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
AES interacts with corepressor proteins to form a complex that modulates transcriptional activity. This complex involvement is critical for developmental processes and cell differentiation affecting how cells acquire specialized functions. Through these interactions AES influences the activities of target genes that are essential for organismal development. The presence of AES in significant developmental pathways highlights its regulatory function in cellular mechanisms.
Pathways
AES plays an important role in the Notch signaling and Wnt signaling pathways both integral to cellular differentiation and proliferation. AES specifically interacts with components of these pathways acting to modulate their signaling outcomes. In the Notch signaling pathway AES functions by repressing gene expression therefore maintaining proper cellular balance. Similarly within the Wnt signaling pathway AES interacts with other pathway proteins such as TCF7L2 to adjust gene transcription ensuring normal cellular development and proliferation.
Quality control
STR analysis
CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
Biosafety level
EU: 1 US: 1
Adherent/suspension
Adherent
Gender
Male
Target data
Complementary Products
Primary Antibodies
AB137060
Anti-Amino-terminal enhancer of split/AES antibody [EPR8385]
RabMAb|Recombinant|KO Validated
![Immunocytochemistry/ Immunofluorescence - Anti-Amino-terminal enhancer of split/AES antibody [EPR8385] (AB137060)](https://content.abcam.com/products/images/amino-terminal-enhancer-of-split-aes-antibody-epr8385-ab137060--immunocytochemistry-immunofluorescence-img129135.jpg)
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- 0
Cell Lines & Lysates
AB257818
Human AES (Amino-terminal enhancer of split) knockout HCT116 cell lysate
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Product promise
Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.
For licensing inquiries, please contact partnerships@abcam.com