Human ZAP70 knockout Jurkat cell lysate
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- WB
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='/en-us/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
false
- WB
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='/en-us/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
false
Complementary Products
Reactivity data
Product details
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.
What's included?
Properties and storage information
Gene name
Gene editing type
Gene editing method
Knockout validation
Shipped at conditions
Appropriate short-term storage conditions
Appropriate long-term storage conditions
Supplementary information
This supplementary information is collated from multiple sources and compiled automatically.
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.
Quality control
STR analysis
CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
Biosafety level
EU: 1 US: 1
Adherent/suspension
Suspension
Gender
Male
Product promise
Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.
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