AB274939

Human YWHAE knockout HeLa cell line

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YWHAE KO cell line available to order. KO validated by Next Generation Sequencing, Western blot. Free of charge wild type control available. Knockout achieved by CRISPR/Cas9 X = 2 bp deletion after Lys49 of the WT protein Frameshift = 100%. To order both knockout and wild-type control cells: select '2 x 1000000 Cells/vial'. To order only knockout cells: select '1000000 Cells/vial'.

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Lab

Next Generation Sequencing - Human YWHAE knockout HeLa cell line (AB274939)

2 bp deletion after Lys49 of the WT protein

Lab

Western blot - Human YWHAE knockout HeLa cell line (AB274939)

Western blot : Anti-YWHAE antibody [EPR3918] ab92311 staining at 1/1000 dilution, shown in green; Mouse anti alpha Tubulin ab7291 loading control staining at 1/20000 dilution, shown in magenta. A band was observed at 29 kDa in Wild-type HeLa cell lysates with no signal observed at this size in YWHAE knockout HeLa cell line. To generate this image, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 5% Milk in TBS-0.1 % Tween^® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit 800CW and Goat anti-Mouse 680RD at 1/20,000 dilution.

All lanes:

Western blot - Anti-YWHAE antibody [EPR3918] (<a href='/products/primary-antibodies/ywhae-antibody-epr3918-ab92311'>ab92311</a>) at 1/1000 dilution

Lane 1:

Wild-type HeLa cell lysates at 20 µg

Lane 2:

Western blot - Human YWHAE knockout HeLa cell line (ab274939) at 20 µg

Lane 3:

HEK-293T cell lysates at 20 µg

Lane 4:

A549 cell lysates at 20 µg

Secondary

All lanes:

Goat anti-Rabbit 800CW & Goat anti-Mouse 680RD at 1/20000 dilution

Predicted band size: 29 kDa

Observed band size: 29 kDa

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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

Next Generation Sequencing,Western blot

ノックアウト変異

Knockout achieved by CRISPR/Cas9 X = 2 bp deletion after Lys49 of the WT protein Frameshift = 100%

疾病

Adenocarcinoma

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Reactivity data

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製品の詳細

Recommended control: Human wild-type HeLa cell line (ab271142). Please note a wild-type cell line is not automatically included with a knockout cell line order, if required please add recommended wild-type cell line at no additional cost using the code WILDTYPE-TMTK1.

We will provide viable cells that proliferate on revival.

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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製品内容

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出荷温度及び保存条件

遺伝子名

YWHAE

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

Next Generation Sequencing, Western blot

接合型

Homozygous

出荷温度

Dry Ice

短期保存期間

1-2 weeks

短期保存温度

-196°C

長期保存温度

-196°C

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補足情報

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

The YWHAE protein also known as 14-3-3E acts as a regulatory molecule by binding to various phosphoserine-containing proteins. It plays key roles in cellular signaling and exists as a dimer. YWHAE has a molecular mass of around 29 kDa and is widely expressed in tissues including the brain heart and skeletal muscle. Its interactions often rely on its abundant presence in various cell types showing its versatility.

Biological function summary

YWHAE protein interacts with several enzymes and receptors forming complexes that affect their functional states. It is a member of the 14-3-3 family which facilitates signal transduction by binding to phosphorylated serine/threonine residues. This protein influences processes like cell cycle control apoptosis and stress responses emphasizing its significant roles in maintaining cellular homeostasis.

Pathways

YWHAE plays significant roles in the Akt and MAPK pathways. In the Akt pathway YWHAE partners with proteins like BAD and FOXO which are important in controlling apoptosis and cell survival. In the MAPK pathway it regulates multiple proteins including Raf and MEK impacting cell growth and differentiation. Understanding YWHAE's interactions in these pathways highlights its influence in critical signaling events.

YWHAE has been tied to schizophrenia and neurodevelopmental disorders. In schizophrenia it interacts with proteins like DISC1 affecting neuronal migration and synaptic function. In neurodevelopmental disorders alterations in YWHAE can lead to compromised signaling that disrupts normal brain function. These connections highlight the importance of YWHAE in maintaining neurological health.

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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

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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製品プロトコール

Visit the General protocols
Visit the Troubleshooting

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Abcam product promise

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Human YWHAE knockout HeLa cell line