AB264963

Human TRIM24 knockout HeLa cell line

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TRIM24 KO cell line available to order. KO validated. Free of charge wild type control available. Knockout achieved by using CRISPR/Cas9, 2 bp insertion in exon 1 and Insertion of the selection cassette in exon 1. To order both knockout and wild-type control cells: select '2 x 1000000 Cells/vial'. To order only knockout cells: select '1000000 Cells/vial'.

別名を表示する

E3 ubiquitin-protein ligase Trim24, PTC6, RING finger protein 82, RNF82, TF1A, TIF1, TIF1-alpha, TIF1A_HUMAN, Transcription intermediary factor 1-alpha, Transcriptional intermediary factor 1, Transcriptional intermediary factor 1 alpha, Tripartite motif containing 24, Tripartite motif-containing protein 24, hTIF1

3 Images

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Western blot - Human TRIM24 knockout HeLa cell line (AB264963)

Lanes 1- 2 : Merged signal (red and green). Green - ab256491 observed at 140 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.

ab256491 was shown to react with Tripartite Motif Containing 24 in wild-type HeLa cells in western blot. The band observed in 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 HeLa and TRIM24 knockout HeLa cell lysates 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 a 1 in 1000 dilution and a 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='/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 (<a href='/products/cell-lysates/human-trim24-knockout-hela-cell-lysate-ab258246'>ab258246</a>)

Lane 2:

Western blot - Human TRIM24 knockout HeLa cell line (ab264963)

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 line (AB264963)

Sanger seq

Unknown

Sanger Sequencing - Human TRIM24 knockout HeLa cell line (AB264963)

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

Sanger seq

Unknown

Sanger Sequencing - Human TRIM24 knockout HeLa cell line (AB264963)

Allele-1 : 2 bp insertion in exon 1.

Key facts

細胞タイプ

HeLa

生物種

Human

組織

Cervix

製品の状態

Liquid

form

ノックアウト検証方法

Sanger Sequencing

ノックアウト変異

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

疾病

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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製品の詳細

We will provide viable cells that proliferate on revival.

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

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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製品内容

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出荷温度及び保存条件

遺伝子名

TRIM24

遺伝子編集のタイプ

Knockout

遺伝子編集の方法

CRISPR technology

ノックアウト検証方法

Sanger Sequencing

出荷温度

Dry Ice

短期保存温度

-196°C

長期保存温度

-196°C

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取り扱い方法

初回取り扱いガイドライン

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.

継代培養ガイドライン

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.

培養培地

DMEM (High Glucose) + 10% FBS

凍結保存培地

Cell Freezing Medium-DMSO Serum free media, contains 8.7% DMSO in MEM supplemented with methyl cellulose.

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補足情報

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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製品プロトコール

Visit the General protocols
Visit the Troubleshooting

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ターゲットの情報

See full target information TRIM24

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Human TRIM24 knockout HeLa cell line