AB258365

Human CLOCK (KAT13D) knockout HeLa cell lysate

Name: Human CLOCK (KAT13D) knockout HeLa cell lysate (ab258365) | Abcam
Brand: Abcam Limited
SKU: AB258365
Price: 111000 JPY
Availability: InStock

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CLOCK KO cell lysate available now. KO validated. Free of charge wild type control included. Knockout achieved by using CRISPR/Cas9, 1 bp insertion in exon11.

別名を表示する

CLOCK_HUMAN, Circadian Locomotor Output Cycles Kaput, Circadian locomoter output cycles kaput protein, Circadian locomoter output cycles protein kaput, Circadium Locomotor Output Cycles Kaput, Class E basic helix-loop-helix protein 8, Clock circadian regulator, Clock homolog, Clock protein, KIAA0334, bHLHe8, hCLOCK

2 Images

Lab

Western blot - Human CLOCK (KAT13D) knockout HeLa cell lysate (AB258365)

Lane 1 : Wild-type HeLa cell lysate 20 μg

Lane 2 : CLOCK knockout HeLa cell lysate 20 μg

Lane 3 : A431 cell lysate 20 μg

Lane 4 : U-251 MG cell lysate 20 μg

False colour image of Western blot : Anti-KAT13D / CLOCK antibody staining at 1/2000 dilution, shown in green; Mouse anti-Alpha Tubulin [DM1A] (ab7291) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab93804 was shown to bind specifically to KAT13D / CLOCK. A band was observed at 100 kDa in wild-type HeLa cell lysates with no signal observed at this size in CLOCK knockout cell line ab266054 (knockout cell lysate ab258365). The band observed in the knockout lysate lane below 100 kDa is likely to represent a truncated form of KAT13D / CLOCK. This has not been investigated further and the functional properties of the gene product have not been determined. To generate this image, wild-type and CLOCK knockout HeLa cell lysates were analysed.First, 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 IgG H&L (IRDye^® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preabsorbed (ab216776) at 1/20000 dilution.

All lanes:

Western blot - Anti-KAT13D / CLOCK antibody (<a href='/products/primary-antibodies/kat13d-clock-antibody-ab93804'>ab93804</a>) at 1/2000 dilution

Lane 1:

Wild-type HeLa cell lysate at 20 µg

Lane 2:

CLOCK knockout HeLa cell lysate at 20 µg

Lane 2:

Western blot - Human CLOCK (KAT13D) knockout HeLa cell line (<a href='/products/cell-lines/human-clock-kat13d-knockout-hela-cell-line-ab266054'>ab266054</a>)

Lane 3:

A431 cell lysate at 20 µg

Lane 4:

U-251 MG cell lysate at 20 µg

Predicted band size: 95 kDa

Observed band size: 100 kDa

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Sanger Sequencing - Human CLOCK (KAT13D) knockout HeLa cell lysate (AB258365)

Sanger seq

Unknown

Sanger Sequencing - Human CLOCK (KAT13D) knockout HeLa cell lysate (AB258365)

Homozygous : 1 bp insertion in exon11

Key facts

細胞タイプ

HeLa

生物種

Human

組織

Cervix

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, 1 bp insertion in exon11.

疾病

Adenocarcinoma

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Reactivity data

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製品の詳細

Knockout cell lysate achieved by CRISPR/Cas9.

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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製品内容

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出荷温度及び保存条件

遺伝子名

CLOCK

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

Ambient - Can Ship with Ice

短期保存温度

-20°C

長期保存温度

-20°C

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補足情報

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

KAT13D also known as CLOCK is a gene coding for a protein weighing approximately 97 kDa. The CLOCK protein mainly functions as a transcription factor with histone acetyltransferase activity hence its involvement in chromatin remodeling. This protein is highly expressed in the suprachiasmatic nucleus of the brain pancreas and heart. It regulates expression of genes through folding DNA and influencing transcriptional activity playing a significant role in maintaining circadian rhythms. Scientists often use phrases such as 'anti-CLOCK' 'anticlock' or 'anti-clock' when studying its mechanisms as these highlight the protein's regulatory role.

Biological function summary

The CLOCK protein acts as an important component of the circadian rhythm machinery. It forms a heterodimer complex with BMAL1 which activates transcription of other core clock genes. This process drives the rhythmic expression of various genes essential for physiological and behavioral rhythms. Through this function CLOCK influences the timing of many body systems such as sleep-wake cycles feeding and metabolism. By doing so it sets a steady rhythm to coordinate bodily processes with environmental light-dark cycles ensuring optimal biological activity during appropriate times of the day.

Pathways

The CLOCK protein plays an important role in the circadian signaling pathway where its function involves intricate feedback loops. It controls the oscillation of gene expression alongside other clock proteins like PER and CRY. This feedback mechanism is part of the circadian rhythm regulation pathway which directly influences processes such as hormone regulation and cell cycle progression. CLOCK’s relationship with BMAL1 PER and CRY in these pathways highlights its indispensable role in maintaining the synchronization of endogenous biological rhythms with external time cues.

Disruption of the CLOCK gene is associated with diseases such as sleep disorders and mood disorders. Alterations in CLOCK function can lead to irregular sleep patterns such as in the case of delayed sleep phase disorder owing to its role in the circadian timing system. Moreover irregular rhythms in CLOCK expression have been linked to mood disorders like bipolar disorder. The association between CLOCK dysfunction and these disorders highlights its importance alongside its interaction with proteins like CRY and PER in maintaining mental health stability.

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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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Abcam product promise

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Human CLOCK (KAT13D) knockout HeLa cell lysate