AB273795

Human ZAP70 knockout Jurkat cell lysate

Name: Human ZAP70 knockout Jurkat cell lysate (ab273795) | Abcam
Brand: Abcam Limited
SKU: AB273795
Price: 107000 JPY
Availability: InStock

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ZAP70 KO cell lysate available now. KO validated by Next Generation Sequencing. Free of charge wild type control included. Knockout achieved by CRISPR/Cas9 X = 2 bp insertion Frameshift: 100%.

別名を表示する

70 kDa zeta-chain associated protein, Tyrosine-protein kinase ZAP-70, SRK, Syk-related tyrosine kinase, ZAP70

2 Images

Lab

Western blot - Human ZAP70 knockout Jurkat cell lysate (AB273795)

Lane 1 : Wild-type Jurkat cell lysate 20 μg
Lane 2 : ZAP70 knockout Jurkat cell lysate 20 μg
Lane 3 : MOLT-4 cell lysate 20 μg
Lane 4 : Raji cell lysate 20 μg

False colour image of Western blot : Anti-ZAP70 antibody [E267] staining at 1/500 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab32410 was shown to bind specifically to ZAP70. A band was observed at 70 kDa in wild-type Jurkat cell lysates with no signal observed at this size in ZAP70 knockout cell line ab273841 (knockout cell lysate ab273795). The band observed in the knockout lysate lane below 70 kDa is likely to represent a truncated form of ZAP70. 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 ZAP70 knockout Jurkat cell lysates were analysed.First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % 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-ZAP70 antibody [E267] (<a href='/products/primary-antibodies/zap70-antibody-e267-ab32410'>ab32410</a>) at 1/500 dilution

Lane 1:

Wild-type Jurkat cell lysate at 20 µg

Lane 2:

ZAP70 knockout Jurkat cell lysate at 20 µg

Lane 3:

MOLT-4 cell lysate at 20 µg

Lane 4:

Raji cell lysate at 20 µg

Predicted band size: 69 kDa

Observed band size: 70 kDa

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Lab

Western blot - Human ZAP70 knockout Jurkat cell lysate (AB273795)

Lane 1 : Wild-type Jurkat cell lysate 20 μg
Lane 2 : ZAP70 knockout Jurkat cell lysate 20 μg
Lane 3 : MOLT-4 cell lysate 20 μg
Lane 4 : Raji cell lysate 20 μg

False colour image of Western blot : Anti-ZAP70 antibody [YE291] staining at 1/500 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab32429 was shown to bind specifically to ZAP70. A band was observed at 70 kDa in wild-type Jurkat cell lysates with no signal observed at this size in ZAP70 knockout cell line ab273841 (knockout cell lysate ab273795). The band observed in the knockout lysate lane below 70 kDa is likely to represent a truncated form of ZAP70. 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 ZAP70 knockout Jurkat cell lysates were analysed.First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % 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-ZAP70 antibody [YE291] (<a href='/products/primary-antibodies/zap70-antibody-ye291-ab32429'>ab32429</a>) at 1/500 dilution

Lane 1:

Wild-type Jurkat cell lysate at 20 µg

Lane 2:

ZAP70 knockout Jurkat cell lysate at 20 µg

Lane 3:

MOLT-4 cell lysate at 20 µg

Lane 4:

Raji cell lysate at 20 µg

Predicted band size: 69 kDa

Observed band size: 70 kDa

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Key facts

細胞タイプ

Jurkat

生物種

Human

組織

Blood

ノックアウト検証方法

Next Generation Sequencing

ノックアウト変異

Knockout achieved by CRISPR/Cas9 X = 2 bp insertion Frameshift: 100%

疾病

Non-Hodgkin Lymphoma

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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>Western blot data indicates that the CRISPR gene edit may have resulted in a truncation of the protein of interest. Please see data images.</p>" } } }

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製品の詳細

Western blot data indicates that the CRISPR gene edit may have resulted in a truncation of the protein of interest. Please see data images.

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

{ "values": { "1Kit": { "sellingSize": "1 Kit", "publicAssetCode":"ab273795-1Kit", "assetComponentDetails": [ { "size":"1 x 100 µg", "name":"Human wild-type Jurkat cell lysate", "number":"AB273795-CMP02", "productcode":"" }, { "size":"1 x 100 µg", "name":"Human ZAP70 knockout Jurkat cell lysate", "number":"AB273795-CMP01", "productcode":"" } ] } } }

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出荷温度及び保存条件

遺伝子名

ZAP70

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

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.

ZAP70 also known as Zeta-chain-associated protein kinase 70 is a tyrosine kinase that plays an important role in T-cell receptor signaling. The protein has a molecular mass of approximately 70 kDa. ZAP70 is mainly expressed in T cells and natural killer (NK) cells. It gets activated upon T-cell receptor engagement initiating downstream signaling cascades essential for T-cell function and adaptive immune response.

Biological function summary

ZAP70 is instrumental in the signaling cascade that activates T cells in response to antigen recognition. It forms part of a larger signaling complex following the engagement of the T-cell receptor with antigens. This complex transmits critical activation signals leading to further cellular responses such as cytokine production and cell proliferation. ZAP70's function ensures appropriate immune responses and helps maintain immune system balance.

Pathways

ZAP70 plays an integral role in T-cell receptor signaling pathways. This protein interacts with other signaling molecules like Lck and LAT facilitating the transmission of activation signals within the cell. As part of the T-cell activation pathway ZAP70 helps to bridge receptor-ligand interactions with cellular responses enabling the immune cells to combat infections effectively.

ZAP70 mutations or dysregulations can lead to severe immunodeficiencies or autoimmune diseases. For example ZAP70 deficiency results in severe combined immunodeficiency (SCID) which severely impairs immune function. Additionally aberrant ZAP70 activity has links to certain autoimmune disorders like rheumatoid arthritis where it influences abnormal immune signaling. Understanding the biochemical pathways that involve ZAP70 provides insight into managing these diseases and developing therapeutic strategies.

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Quality control

STR analysis

CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX

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Cell culture

Biosafety level

EU: 1 US: 1

Adherent/suspension

Suspension

Gender

Male

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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 ZAP70 knockout Jurkat cell lysate