AB257131

Human ADAR (ADAR1) knockout HEK-293T cell lysate

Name: Human ADAR (ADAR1) knockout HEK-293T cell lysate (ab257131) | Abcam
Brand: Abcam Limited
SKU: AB257131
Price: 119000 JPY
Availability: InStock

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ADAR KO cell lysate available now. KO validated by Western blot. Free of charge wild type control included. Knockout achieved by using CRISPR/Cas9, 17 bp deletion in exon2 and 1 bp insertion in exon2.

別名を表示する

136 kDa double-stranded RNA-binding protein, ADAR, AGS6, AV242451, Adenosine deaminase RNA specific, Adenosine deaminase RNA specific 1, Adenosine deaminase acting on RNA 1 A, Adenosine deaminase that act on RNA, DRADA, DSRAD_HUMAN, Double-stranded RNA-specific adenosine deaminase, Double-stranded RNA-specific editase Adar, Dsh, EC 3.5.4.-, G1P1, Ifi4 protein, Interferon induced protein 4, Interferon-inducible protein 4, K88DSRBP, P136, Pre-mRNA adenosine deaminase, RNA adenosine deaminase 1, RNA-editing deaminase 1, RNA-editing enzyme 1, dsRNA adenosine deaminase, mZaADAR

4 Images

Lab

Western blot - Human ADAR (ADAR1) knockout HEK-293T cell lysate (AB257131)

Lane 1 : Wild-type HEK293T cell lysate (20 ug)
Lane 2 : ADAR knockout HEK293T cell lysate (20 ug)
Lane 3 : HeLa cell lysate (20 ug)

ab126745 was shown to specifically react with ADAR1 in wild-type HEK293T cells. Loss of signal was observed when knockout cell line ab266846 (knockout cell lysate ab257131) was used. Wild-type and ADAR1 knockout samples were subjected to SDS-PAGE. ab126745 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4^oC 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-ADAR1 antibody [EPR7033] (<a href='/products/primary-antibodies/adar1-antibody-epr7033-ab126745'>ab126745</a>) at 1/1000 dilution

Lane 1:

Wild-type HEK293T cell lysate at 20 µg

Lane 2:

ADAR knockout HEK293T cell lysate at 20 µg

Lane 2:

Western blot - Human ADAR (ADAR1) knockout HEK-293T cell line (<a href='/products/cell-lines/human-adar-adar1-knockout-hek-293t-cell-line-ab266846'>ab266846</a>)

Lane 3:

HeLa 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: 136 kDa,19 kDa,29 kDa,35 kDa,36 kDa

Observed band size: 130 kDa,20 kDa,29 kDa,35 kDa,36 kDa

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Supplier Data

Western blot - Human ADAR (ADAR1) knockout HEK-293T cell lysate (AB257131)

Blocking and diluting buffer and concentration : 5% NFDM/TBST Performed under reducing conditions. In Western blot, ab307585 was shown to bind specifically to ADAR1. A band was observed at 150 kDa in wild-type HEK-293T cell lysates whereas no signal observed at this size in ADAR1 knockout cell line ab266846 (knockout cell lysate ab257131). Exposure time : 59 seconds

All lanes:

Western blot - Anti-ADAR1 antibody [EPR25431-60] (<a href='/products/primary-antibodies/adar1-antibody-epr25431-60-ab307585'>ab307585</a>) at 1/1000 dilution

Lane 1:

Wild-type HEK-293T (human embryonic kidney epithelial cell) whole cell lysate at 20 µg

Lane 2:

ADAR1 knockout HEK-293T whole cell lysate at 20 µg

Lane 2:

Western blot - Human ADAR (ADAR1) knockout HEK-293T cell line (<a href='/products/cell-lines/human-adar-adar1-knockout-hek-293t-cell-line-ab266846'>ab266846</a>)

Lane 2:

Western blot - Human ADAR (ADAR1) knockout HEK-293T cell lysate (ab257131)

Lane 3:

HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lane 4:

Ramos (human burkitts lymphoma b lymphocyte) whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Observed band size: 150 kDa

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Exposure time: 59s

Sanger Sequencing - Human ADAR (ADAR1) knockout HEK-293T cell lysate (AB257131)

Sanger seq

Unknown

Sanger Sequencing - Human ADAR (ADAR1) knockout HEK-293T cell lysate (AB257131)

Allele-2 : 1 bp insertion in exon2

Sanger seq

Unknown

Sanger Sequencing - Human ADAR (ADAR1) knockout HEK-293T cell lysate (AB257131)

Allele-1 : 17 bp deletion in exon2

Key facts

細胞タイプ

HEK-293T

生物種

Human

組織

Kidney

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, 17 bp deletion in exon2 and 1 bp insertion in exon2.

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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

ADAR

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

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.

ADAR1 also known as RNA-specific adenosine deaminase 1 or ADAR is an enzyme with a mass of approximately 150 kDa. This protein targets double-stranded RNA (dsRNA) and acts mechanistically to convert adenosine to inosine in pre-mRNA sequences a process known as A-to-I RNA editing. ADAR1 is expressed in numerous tissues with high levels in the brain liver and lungs. Its localization within cells can vary often found in both the nucleus and cytoplasm which influences its function.

Biological function summary

ADAR1 plays a role in the regulation of RNA molecules affecting their stability and translation. It is linked to the editosome complex working alongside other proteins to perform RNA editing tasks. By modifying the coding potential of mRNAs ADAR1 contributes to the diversity of proteomes and helps manage the responses to viral RNAs giving the immune system tools to recognize endogenous and exogenous RNA.

Pathways

ADAR1 is significant in the interferon signaling pathway and RNA processing pathways. It operates in coordination with proteins like PKR which is involved in the response to viral infections. ADAR1 ensures that the immune response is not directed against the self highlighting its role in the regulation of the innate immune system. These pathways are critical in maintaining homeostasis and preventing unchecked immune responses.

Mutations or dysregulation of ADAR1 have associations with autoimmune diseases like Aicardi-Goutieres syndrome and certain cancers. The enzyme's role in editing RNA makes it essential in preventing inappropriate immune attacks against the body's own cells highlighting its interaction with MDA5 another protein involved in immune regulation. Understanding ADAR1 and its related pathways may offer potential therapeutic targets for these conditions including exploration into ADAR1 inhibitors as interventions.

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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 ADAR (ADAR1) knockout HEK-293T cell lysate