AB265793

Human TIAL1 knockout HeLa cell line

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TIAL1 KO cell line available to order. KO validated by Western blot. Free of charge wild type control available. Knockout achieved by using CRISPR/Cas9, 1 bp insertion in exon 2 and Insertion of the selection cassette in exon 2. 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

Aging associated gene 7 protein, Nucleolysin TIAR, T cluster binding protein, TCBP, TIA 1 related nucleolysin, TIA-1-related protein, TIA1 cytotoxic granule associated RNA binding protein like 1, TIAR_HUMAN

4 Images

Lab

Western blot - Human TIAL1 knockout HeLa cell line (AB265793)

Lanes 1-4 : Merged signal (red and green). Green - ab169547 observed at 49 kDa. Red - loading control ab8245 observed at 36 kDa.

ab169547 Anti-TIAL1 antibody [EPR11323(B)] was shown to specifically react with TIAL1 in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab265793 (knockout cell lysate ab257743) was used. Wild-type and TIAL1 knockout samples were subjected to SDS-PAGE. ab169547 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-TIAL1 antibody [EPR11323(B)] (<a href='/products/primary-antibodies/tial1-antibody-epr11323b-ab169547'>ab169547</a>) at 1/1000 dilution

Lane 1:

Wild-type HeLa cell lysate at 20 µg

Lane 2:

TIAL1 knockout HeLa cell lysate at 20 µg

Lane 2:

Western blot - Human TIAL1 knockout HeLa cell line (ab265793)

Lane 3:

Jurkat cell lysate at 20 µg

Lane 4:

K-562 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 1/10000 dilution

Predicted band size: 41 kDa

Observed band size: 49 kDa

false

Sanger Sequencing - Human TIAL1 knockout HeLa cell line (AB265793)

Sanger seq

Unknown

Sanger Sequencing - Human TIAL1 knockout HeLa cell line (AB265793)

Allele-1 : 1 bp insertion in exon 2.

Sanger seq

Unknown

Sanger Sequencing - Human TIAL1 knockout HeLa cell line (AB265793)

Allele-3 : Insertion of the selection cassette in exon 2.

Sanger seq

Unknown

Sanger Sequencing - Human TIAL1 knockout HeLa cell line (AB265793)

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

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, 1 bp insertion in exon 2 and Insertion of the selection cassette in exon 2

疾病

Adenocarcinoma

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Reactivity data

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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

TIAL1

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

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.

TIAL1 also known as TIA-1 related protein is a 42 kDa protein that plays a mechanical role in RNA processing. It belongs to the RNA-binding protein family and contains three RNA recognition motifs and a glutamine-rich domain. TIAL1 is widely expressed in various tissues including the spleen thymus and brain. It influences the regulation of mRNA stability and translation specifically recognizing and binding to U-rich elements in the 3'-untranslated regions of target mRNAs.

Biological function summary

TIAL1 influences gene expression by associating with stress granules cytoplasmic aggregates formed in response to cellular stress. This association helps inhibit translation initiation modulating the cellular stress response. TIAL1 is part of a complex with other RNA-binding proteins that integrate signaling events and manage mRNA turnover and stress granule dynamics. Through this interaction it affects apoptosis and contributes to cell survival under stress conditions.

Pathways

Studies demonstrate that TIAL1 plays a role in the apoptotic signaling pathway and the stress response pathway. By interacting with proteins such as eIF2α kinase TIAL1 helps mediate the cellular response to environmental and oxidative stress. These pathways maintain cellular homeostasis and TIAL1 assists in regulating translation during stress ensuring proper cell function.

TIAL1 has relevance in neurodegenerative diseases like Alzheimer's and amyotrophic lateral sclerosis (ALS). The formation of stress granules influenced by TIAL1 is connected to the pathological misfolding of proteins implicated by related proteins such as TDP-43 and FUS. Faulty regulation of these processes may lead to the progression of neurodegenerative conditions highlighting TIAL1's significant role in their development.

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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 Nucleolysin TIAR

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Human TIAL1 knockout HeLa cell line