AB258246

Human TRIM24 knockout HeLa cell lysate

Name: Human TRIM24 knockout HeLa cell lysate (ab258246) | Abcam
Brand: Abcam Limited
SKU: AB258246
Price: 645 USD
Availability: InStock

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TRIM24 KO cell lysate available now. KO validated. Free of charge wild type control included. Knockout achieved by using CRISPR/Cas9, 2 bp insertion in exon 1 and Insertion of the selection cassette in exon 1.

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Western blot - Human TRIM24 knockout HeLa cell lysate (AB258246)

Lane 1 : Wild-type HeLa cell lysate (20μg)

Lane 2 : TRIM24 knockout HeLa cell lysate (20μg)

Lane 3 : Hap1 cell lysate (20μg)

Lane 4 : A549 cell lysate (20μg)

Lanes 1- 4 : Merged signal (red and green). Green - ab256491 observed at 140 kDa. Red - loading control, ab8245 observed at 37 kDa.

ab256491 Anti-TRIM24 antibody [EPR22825-2] was shown to specifically react with Tripartite Motif Containing 24 in wild-type HeLa cells in western blot. The band observed in the knockout cell line ab264963 (knockout cell lysate ab258246) lane below 140kDa may represent truncated forms and cleaved fragments. This has not been investigated further. Wild-type and Tripartite Motif Containing 24 knockout samples were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab256491 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4^°C at 1 in 1000 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-TRIM24 antibody [EPR22825-2] (<a href='/en-us/products/primary-antibodies/trim24-antibody-epr22825-2-ab256491'>ab256491</a>) at 1/1000 dilution

Lane 1:

Wild-type HeLa cell lysate at 20 µg

Lane 2:

TRIM24 knockout HeLa cell lysate at 20 µg

Lane 2:

Western blot - Human TRIM24 knockout HeLa cell lysate (ab258246)

Lane 2:

Western blot - Human TRIM24 knockout HeLa cell line (<a href='/en-us/products/cell-lines/human-trim24-knockout-hela-cell-line-ab264963'>ab264963</a>)

Lane 3:

Hap1 cell lysate at 20 µg

Lane 4:

A549 cell lysate at 20 µg

Predicted band size: 116 kDa

Observed band size: 140 kDa

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Sanger Sequencing - Human TRIM24 knockout HeLa cell lysate (AB258246)

Sanger seq

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Sanger Sequencing - Human TRIM24 knockout HeLa cell lysate (AB258246)

Allele-2 : Insertion of the selection cassette in exon 1

Sanger seq

Unknown

Sanger Sequencing - Human TRIM24 knockout HeLa cell lysate (AB258246)

Allele-1 : 2 bp insertion in exon 1

Key facts

Cell type

HeLa

Species or organism

Human

Tissue

Cervix

Knockout validation

Sanger Sequencing

Mutation description

Knockout achieved by using CRISPR/Cas9, 2 bp insertion in exon 1 and Insertion of the selection cassette in exon 1.

Disease

Adenocarcinoma

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Reactivity data

{ "title": "Reactivity Data", "filters": { "stats": ["", "Reactivity", "Dilution Info", "Notes"] }, "values": { "WB": { "reactivity":"TESTED_AND_REACTS", "dilution-info":"", "notes":"<p>Western blot data indicates that the CRISPR gene edit may have resulted in a truncation of the protein of interest. Please see data images.</p>" } } }

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Product details

Western blot data indicates that the CRISPR gene edit may have resulted in a truncation of the protein of interest. Please see data images.

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, ERS Genomics Limited and Sigma-Aldrich Co. LLC, and is developed with patented technology. For full details of the licenses and patents please refer to our limited use license and patent pages.

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What's included?

{ "values": { "1Kit": { "sellingSize": "1 Kit", "publicAssetCode":"ab258246-1Kit", "assetComponentDetails": [ { "size":"1 x 100 µg", "name":"Human TRIM24 knockout HeLa cell lysate", "number":"AB258246-CMP02", "productcode":"" }, { "size":"1 x 100 µg", "name":"Human wild-type HeLa cell lysate", "number":"AB258246-CMP01", "productcode":"" } ] } } }

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Properties and storage information

Gene name

TRIM24

Gene editing type

Knockout

Gene editing method

CRISPR technology

Knockout validation

Sanger Sequencing

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.

TRIM24 also known as transcription intermediary factor 1-alpha (TIF1α) is a protein that acts mechanically as a transcriptional regulator. It has a molecular mass of approximately 140 kDa. This protein contains several domains including a tripartite motif (the eponymous TRIM) which consists of a RING domain B-box domains and a coiled-coil region. TRIM24 interacts with nuclear receptors and is expressed in various tissues but shows higher expression in the liver and lungs. It also plays a role in bridging the interaction between chromatin and transcription factors.

Biological function summary

TRIM24 is involved in transcriptional regulation by influencing gene expression. It acts as a co-regulator being part of large protein complexes. This protein interacts directly with histone tails to read histone marks and release transcriptional repression. TRIM24 also plays a role in ubiquitination and subsequent proteasomal degradation of specific proteins suggesting its involvement in maintaining protein homeostasis. Additionally TRIM24 has been shown to interact with p53 influencing cell cycle regulation and apoptosis.

Pathways

TRIM24 is involved in the regulation of the retinoic acid and vitamin D signaling pathways. It modulates the transcriptional activity of nuclear receptors through these pathways influencing cell proliferation and differentiation processes. TRIM24 does so by interacting with other proteins such as retinoid X receptor (RXR) and estrogen receptor (ER) establishing important crosstalk between signaling cascades critical for cell fate decisions.

TRIM24 has been implicated in the development of certain cancers such as breast cancer and liver cancer. In these contexts it functions as an oncogene promoting tumorigenesis through aberrant transcriptional regulation. TRIM24 can interact with other proteins associated with oncogenesis including p53 modulating its tumor suppressor functions. Furthermore deregulation of TRIM24 activity or expression levels has been correlated with inflammatory disorders highlighting its relevance in both cancer and inflammation-related conditions.

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Quality control

STR analysis

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

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Cell culture

Biosafety level

EU: 2 US: 2

Adherent/suspension

Adherent

Gender

Female

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Product protocols

Visit the General protocols
Visit the Troubleshooting

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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

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

Human TRIM24 knockout HeLa cell lysate

Western blot - Human TRIM24 knockout HeLa cell lysate (AB258246)

Sanger Sequencing - Human TRIM24 knockout HeLa cell lysate (AB258246)

Sanger Sequencing - Human TRIM24 knockout HeLa cell lysate (AB258246)

Key facts

Cell type

Species or organism

Tissue

Knockout validation

Mutation description

Disease

Complementary Products

Primary Antibodies

Primary Antibodies

Primary Antibodies

Primary Antibodies

Reactivity data

Product details

What's included?

Properties and storage information

Gene name

Gene editing type

Gene editing method

Knockout validation

Shipped at conditions

Appropriate short-term storage conditions

Appropriate long-term storage conditions

Supplementary information

Biological function summary

Pathways

Quality control

STR analysis

Cell culture

Biosafety level

Adherent/suspension

Gender

Product protocols

Product promise