Human ZAP70 knockout Jurkat cell line
- Advanced Validation
- What is this?
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- WB
Lab
Western blot - Human ZAP70 knockout Jurkat cell line (AB273841)
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 CRISPR-Cas9 edited cell line ab273841 (CRISPR-Cas9 edited cell lysate ab273795). The band observed in the CRISPR-Cas9 edited 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 CRISPR-Cas9 edited 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 °. 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 CRISPR-Cas9 edited Jurkat cell lysate at 20 µg
Lane 2:
Western blot - Human ZAP70 knockout Jurkat cell line (ab273841)
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 line (AB273841)
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 CRISPR-Cas9 edited cell line ab273841 (CRISPR-Cas9 edited cell lysate ab273795). The band observed in the CRISPR-Cas9 edited 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 CRISPR-Cas9 edited 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 °. 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 CRISPR-Cas9 edited Jurkat cell lysate at 20 µg
Lane 2:
Western blot - Human ZAP70 knockout Jurkat cell line (ab273841)
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
- NGS
Lab
Next Generation Sequencing - Human ZAP70 knockout Jurkat cell line (AB273841)
2 bp insertion after Pro90 of the WT protein
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Cell Lines & Lysates
AB266952
Human ZC3HAV1 (Zinc finger antiviral protein) knockout A549 cell line
- 0
- 0
Reactivity data
Product details
We will provide viable cells that proliferate on revival.
Western blot data indicates that the CRISPR gene edit may have resulted in a truncation of the protein of interest. Please see data images.
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
Handling procedures
Initial handling guidelines
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 for bath 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. Based on cell count, seed cells in an appropriate cell culture flask at a density of 2x105 cells/mL. Seeding density is given as a guide only and should be scaled to align with individual lab schedules.
4. Incubate the culture at 37°C incubator with 5% CO2. Cultures should be monitored daily.
Subculture guidelines
- All seeding densities should be based on cell counts gained by established methods.
- A guide seeding density of 2x105 cells/mL is recommended.
- Do not allow cell density to exceed 3x106 cells/mL.
Culture medium
RPMI + 10% FBS
Cryopreservation medium
Cell Freezing Medium-DMSO Serum free media, contains 8.7% DMSO in MEM supplemented with methyl cellulose.
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
Target data
Product promise
Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.
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