Human SYK knockout HEK-293T cell line
- Advanced Validation
- What is this?
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SYK KO cell line available to order. KO validated by Western blot. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, Homozygous: 127 bp deletion in exon 2.
View Alternative Names
EC 2.7.10.2, KSYK_HUMAN, Spleen tyrosine kinase, Tyrosine-protein kinase SYK, kinase Syk, p72-Syk
- WB
Lab
Western blot - Human SYK knockout HEK-293T cell line (AB282649)
False colour image of Western blot : Anti-Syk antibody [SYK-01] staining at 1 ug/ml shown in green; Rabbit Anti-GAPDH antibody [EPR16891] (ab181602) loading control staining at 1/20000 dilution shown in red. In Western blot ab3993 was shown to bind specifically to Syk. A band was observed at 72/73 kDa in wild-type HEK-293T cell lysates with no signal observed at this size in SYK knockout cell line ab282649 (knockout cell lysate ab283048). To generate this image wild-type and SYK knockout HEK-293T 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-Mouse IgG H&L (IRDye® 800CW) preabsorbed (ab216772) and Goat anti-Rabbit IgG H&L (IRDye® 680RD) preabsorbed (ab216777) at 1/20000 dilution.
All lanes:
Western blot - Anti-Syk antibody [SYK-01] (<a href='/en-us/products/primary-antibodies/syk-antibody-syk-01-ab3993'>ab3993</a>) at 1 µg/mL
Lane 1:
Wild-type HEK-293T cell lysate at 20 µg
Lane 2:
SYK knockout HEK-293T cell lysate at 20 µg
Lane 2:
Western blot - Human SYK knockout HEK-293T cell line (ab282649)
Lane 3:
HAP1 cell lysate at 20 µg
Predicted band size: 72 kDa
Observed band size: 72 kDa,73 kDa
false
- WB
Lab
Western blot - Human SYK knockout HEK-293T cell line (AB282649)
False colour image of Western blot : Anti-Syk antibody [EP573Y] staining at 1/1000 dilution shown in green; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution shown in red. In Western blot ab40781 was shown to bind specifically to Syk. A band was observed at 70/72 kDa in wild-type HEK-293T cell lysates with no signal observed at this size in SYK knockout cell line ab282649 (knockout cell lysate ab283048). To generate this image wild-type and SYK knockout HEK-293T 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-Syk antibody [EP573Y] (<a href='/en-us/products/primary-antibodies/syk-antibody-ep573y-ab40781'>ab40781</a>) at 1/1000 dilution
Lane 1:
Wild-type HEK-293T cell lysate at 20 µg
Lane 2:
SYK knockout HEK-293T cell lysate at 20 µg
Lane 2:
Western blot - Human SYK knockout HEK-293T cell line (ab282649)
Predicted band size: 72 kDa
Observed band size: 70 kDa
false
- Sanger seq
Supplier Data
Sanger Sequencing - Human SYK knockout HEK-293T cell line (AB282649)
127 bp deletion in exon 2
Reactivity data
Product details
Recommended control: Human wild-type HEK-293T cell line (ab282205). Please note a wild-type cell line is not automatically included with a knockout cell line order, if required please add recommended wild-type cell line at no additional cost using the code WILDTYPE-TMTK1.
We will provide viable cells that proliferate on revival.
This product is subject to limited use licenses from The Broad Institute and ERS Genomics Limited, and is developed with patented technology. For full details of the limited use licenses and relevant 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
Zygosity
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 bath for 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 seed all remaining cells into a T25.
4. Incubate the culture at 37°C incubator with 5% CO2. Check the culture one day after revival and continue to check until 80% confluent. Media change can be given if needed.
5. Once confluent passage into an appropriate flask at a density of 2x104 cells/cm2. Seeding density is given as a guide only and should be scaled to align with individual lab schedules. 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 2x104 cells/cm2 is recommended.
- Cells should be passaged when they have achieved 80-90% confluence.
Culture medium
DMEM (High Glucose) + 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
Syk plays a role in transmitting signals from the cell surface to the interior of the cell facilitating various cellular activities. It is a part of signal transduction complexes and functions in the regulation of immune cell activation differentiation and survival. It affects processes like Fc receptor signaling in macrophages and neutrophils impacting immune responses and inflammation. The phospho-Syk form influences the strength and duration of signaling affecting various cellular functions.
Pathways
Syk is significant in the B-cell receptor (BCR) signaling pathway and Fc receptor signaling pathway. Within these pathways Syk interacts with proteins like Lyn and Fyn which are other kinases involved in BCR signal transduction. Syk phosphorylation noted as p-Syk plays a part in activating downstream molecules such as phospholipase C gamma (PLCγ) to mediate cellular responses to external stimuli. The critical role of Syk in these pathways influences both adaptive and innate immune responses.
Quality control
STR analysis
CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
Biosafety level
EU: 2 US: 2
Adherent/suspension
Adherent
Gender
Female
Target data
Complementary Products
Primary Antibodies
AB62350
Anti-Syk (phospho Y525) antibody [EP575(2)Y]
20ul selling size|RabMAb|Recombinant
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- 0
- 0
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Product promise
Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.
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