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AB257818

Human AES (Amino-terminal enhancer of split) knockout HCT116 cell lysate

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TLE5 KO cell lysate available now. KO validated by Western blot. Free of charge wild type control included. Knockout achieved by using CRISPR/Cas9, Homozygous: 1 bp deletion in exon2.
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Western blot - Human AES (Amino-terminal enhancer of split) knockout HCT116 cell lysate (AB257818)
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Lab

Western blot - Human AES (Amino-terminal enhancer of split) knockout HCT116 cell lysate (AB257818)

Lane 1 : Wild-type HCT 116 (Human colorectal carcinoma cell line) whole cell lysate (20 ug)
Lane 2 : AES knockout HCT 116 (Human colorectal carcinoma cell line) whole cell lysate (20 ug)
Lane 3 : HEK-293T (Human epithelial cell line from embryonic kidney transformed with large T antigen) whole cell lysate (20 ug)
Lane 4 : PC3 (Human prostate adenocarcinoma cell line) whole cell lysate (20 ug)

ab137060 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 4oC 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 (<a href='/en-us/products/cell-lines/human-aes-amino-terminal-enhancer-of-split-knockout-hct116-cell-line-ab266886'>ab266886</a>)

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

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Sanger Sequencing - Human AES (Amino-terminal enhancer of split) knockout HCT116 cell lysate (AB257818)
  • Sanger seq

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Sanger Sequencing - Human AES (Amino-terminal enhancer of split) knockout HCT116 cell lysate (AB257818)

Homozygous : 1 bp deletion in exon2

Key facts

Cell type

HCT116

Species or organism

Human

Tissue

Colon

Knockout validation

Sanger Sequencing,Western blot

Mutation description

Knockout achieved by using CRISPR/Cas9, Homozygous: 1 bp deletion in exon2.

Disease

Carcinoma

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Complementary Products

Primary Antibodies

AB137060

Anti-Amino-terminal enhancer of split/AES antibody [EPR8385]

RabMAb|Recombinant|KO Validated

Western blot - Anti-Amino-terminal enhancer of split/AES antibody [EPR8385] (AB137060)

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Cell Lines & Lysates

AB266886

Human AES (Amino-terminal enhancer of split) knockout HCT116 cell line

Advanced Validation

Western blot - Human AES (Amino-terminal enhancer of split) knockout HCT116 cell line (AB266886)

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Reactivity data

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Product details

Knockout cell lysate achieved by CRISPR/Cas9.

REACH authorisation
Abcam has not and does not intend to apply for the REACH Authorisation of customers' uses of products that contain European Authorisation list (Annex XIV) substances.
It is the responsibility of our customers to check the necessity of application of REACH Authorisation, and any other relevant authorisations, for their intended uses.

Lysate preparation: Our lysates are made using RIPA buffer to which we add a protease inhibitor cocktail and phosphatase inhibitor cocktail (ratio: 300:100:10). This means that the protein of interest is denatured. If you require a native form of the protein please use the live cell version. Please refer to our lysis protocol for further details on how our lysates are prepared.

User storage instructions: Lyophilizate may be stored at 4°C. After reconstitution, store at -20°C for short-term storage or -80°C for long-term storage.

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.

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What's included?

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Properties and storage information

Gene name
TLE5
Gene editing type
Knockout
Gene editing method
CRISPR technology
Knockout validation
Sanger Sequencing, Western blot
Zygosity
Homozygous
Shipped at conditions
Ambient - Can Ship with Ice
Appropriate short-term storage conditions
-20°C
Appropriate long-term storage conditions
-20°C
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Supplementary information

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

The Amino-terminal enhancer of split commonly known as AES is a protein with a molecular mass of approximately 22 kDa. It functions as a transcriptional repressor and is involved in controlling gene expression by interacting with DNA. AES is expressed in various tissues including the brain heart and liver indicating its broad role in different biological processes. Its ability to inhibit specific gene expression highlights its involvement in intricate regulatory mechanisms. Researchers also recognize the protein by other names such as enhancer protein and protein enhancer.
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.

AES has connections to certain types of cancer and neurodevelopmental disorders. Aberrant expression or mutation of AES can lead to disruptions in normal signaling contributing to pathological conditions. In cancer the dysregulation of the Notch signaling pathway where AES plays an important regulatory role is often associated with tumorigenesis. Additionally AES interactions with other proteins like TCF7L2 in Wnt signaling pathways may be implicated in colorectal cancer progression. In neurodevelopmental disorders alterations in the AES function can affect brain development potentially leading to conditions like autism spectrum disorders.
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Quality control

STR analysis

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

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Cell culture

Biosafety level

EU: 1 US: 1

Adherent/suspension

Adherent

Gender

Male

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Product protocols

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Product promise

We are committed to supporting your work with high-quality reagents, and we're here for you every step of the way. In the unlikely event that one of our products does not perform as expected, you're protected by our Product Promise.
For full details, please see our Terms & Conditions

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