AB265878

Human TRIM29 knockout HeLa cell line

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TRIM29 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: 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

ATDC, Ataxia telangiectasia group D-associated protein, FLJ36085, TRI29_HUMAN, Tripartite motif containing 29, Tripartite motif protein 29, Tripartite motif protein TRIM29, Tripartite motif-containing protein 29

2 Images

Lab

Western blot - Human TRIM29 knockout HeLa cell line (AB265878)

Lanes 1-4 : Merged signal (red and green). Green - ab108627 observed at 74 kDa. Red - loading control ab8245 observed at 37 kDa.

ab108627 Anti-TRIM29 antibody [EPR3494] was shown to specifically react with TRIM29 in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab265878 (knockout cell lysate ab257770) was used. Wild-type and TRIM29 knockout samples were subjected to SDS-PAGE. ab108627 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4° 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-TRIM29 antibody [EPR3494] (<a href='/products/primary-antibodies/trim29-antibody-epr3494-ab108627'>ab108627</a>) at 1/1000 dilution

Lane 1:

Wild-type HeLa cell lysate at 20 µg

Lane 2:

TRIM29 knockout HeLa cell lysate at 20 µg

Lane 2:

Western blot - Human TRIM29 knockout HeLa cell line (ab265878)

Lane 3:

A431 cell lysate at 20 µg

Lane 4:

Daudi cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-irdye-800cw-preadsorbed-ab216773'>ab216773</a>) at 1/20000 dilution

Predicted band size: 65 kDa

Observed band size: 74 kDa

false

Sanger Sequencing - Human TRIM29 knockout HeLa cell line (AB265878)

Sanger seq

Unknown

Sanger Sequencing - Human TRIM29 knockout HeLa cell line (AB265878)

Homozygous : Insertion of the selection cassette 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,Western blot

ノックアウト変異

Knockout achieved by using CRISPR/Cas9, Homozygous: 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></p>" } } }

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製品の詳細

We will provide viable cells that proliferate on revival.

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

TRIM29

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, Western blot

接合型

Homozygous

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.

The tripartite motif-containing protein 29 (TRIM29) also known as ATDC is a protein with a molecular mass of approximately 66 kDa. TRIM29 is part of the TRIM family characterized by its RING finger motif B-box type 1 and 2 domains and a coiled-coil region which are significant for its involvement in ubiquitination processes. Researchers have found TRIM29 expression largely in epithelial tissues exhibiting varied levels of expression depending on tissue type.

Biological function summary

TRIM29 functions as an essential regulator of DNA damage responses and cellular proliferation. It is not found as part of any larger protein complex but interacts with various other proteins to carry out its function. TRIM29 can influence the activity of key molecules involved in DNA repair particularly through its interaction with DNA-dependent protein kinase catalytic subunit (DNA-PKcs) which enhances cell survival after genotoxic stress. It participates in ensuring proper functioning of cell cycle checkpoints and modulation of apoptosis.

Pathways

TRIM29 participates in the DNA damage response pathway and the WNT signaling pathway. It interacts with proteins such as β-catenin which influences the transcriptional regulation of target genes involved in cell proliferation. Through these interactions TRIM29 influences cellular responses to DNA damage and the regulation of cell growth linking it to critical pathways that maintain genomic stability and cell integrity.

Aberrant TRIM29 expression correlates with various cancers including breast and colorectal cancers. In breast cancer TRIM29 affects the pathways involving p53 a known tumor suppressor protein. In colorectal cancer its overexpression is associated with enhanced cancer cell proliferation and survival often in conjunction with KRAS mutations. TRIM29's regulatory role in these pathways highlights its potential as a biomarker and a therapeutic target in oncology.

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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 TRIM29

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Human TRIM29 knockout HeLa cell line