Human LCK knockout Jurkat cell line
- Advanced Validation
- What is this?
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LCK KO cell line available to order. KO validated by Next Generation Sequencing, Western blot. Free of charge wild type control provided. Knockout achieved by CRISPR/Cas9; X = 1 bp insertion, 2 bp insertion; Frameshift: 99%.
View Alternative Names
IMD22, LCK proto-oncogene, Src family tyrosine kinase, LCK_HUMAN, LSK, Lck p56, Leukocyte C-terminal Src kinase, Lymphocyte cell-specific protein-tyrosine kinase, Lymphocyte specific protein tyrosine kinase, Membrane associated protein tyrosine kinase, Oncogene lck, Protein YT16, Proto oncogene tyrosine protein kinase LCK, Proto-oncogene Lck, T cell-specific protein-tyrosine kinase, T lymphocyte specific protein tyrosine kinase p56lck, Tyrosine-protein kinase Lck, YT 16, p56(LSTRA) protein tyrosine kinase, p56-LCK, pp58 lck
- WB
Lab
Western blot - Human LCK knockout Jurkat cell line (AB273855)
False colour image of Western blot : Anti-Lck antibody [Y123] 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 ab32149 was shown to bind specifically to Lck. A band was observed at 60 kDa in wild-type Jurkat cell lysates with no signal observed at this size in Lck knockout cell line ab273855 (knockout cell lysate ab273809). To generate this image wild-type and Lck 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 °. 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-Lck antibody [Y123] (<a href='/en-us/products/primary-antibodies/lck-antibody-y123-ab32149'>ab32149</a>) at 1/1000 dilution
Lane 1:
Wild-type Jurkat cell lysate at 20 µg
Lane 2:
Lck knockout Jurkat cell lysate at 20 µg
Lane 2:
Western blot - Human LCK knockout Jurkat cell line (ab273855)
Lane 3:
Ramos cell lysate at 20 µg
Lane 4:
A549 cell lysate at 20 µg
Predicted band size: 58 kDa
Observed band size: 60 kDa
false
- WB
Lab
Western blot - Human LCK knockout Jurkat cell line (AB273855)
False colour image of Western blot : Anti-Lck antibody [EPR20798-107] 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 ab227975 was shown to bind specifically to Lck. A band was observed at 60 kDa in wild-type Jurkat cell lysates with no signal observed at this size in Lck knockout cell line ab273855 (knockout cell lysate ab273809). To generate this image wild-type and Lck 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 °. 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-Lck antibody [EPR20798-107] (<a href='/en-us/products/primary-antibodies/lck-antibody-epr20798-107-ab227975'>ab227975</a>) at 1/1000 dilution
Lane 1:
Wild-type Jurkat cell lysate at 20 µg
Lane 2:
Lck knockout Jurkat cell lysate at 20 µg
Lane 2:
Western blot - Human LCK knockout Jurkat cell line (ab273855)
Lane 3:
Ramos cell lysate at 20 µg
Lane 4:
A549 cell lysate at 20 µg
Predicted band size: 58 kDa
Observed band size: 60 kDa
false
- NGS
Supplier Data
Next Generation Sequencing - Human LCK knockout Jurkat cell line (AB273855)
2 bp insertion (allele 1) and 1 bp insertion (allele 2) after Val65 of the WT protein
- NGS
Supplier Data
Next Generation Sequencing - Human LCK knockout Jurkat cell line (AB273855)
Knockout achieved by CRISPR/Cas9; X = 1 bp insertion, 2 bp insertion; Frameshift : 99%
Reactivity data
Product details
Recommended control: Human wild-type Jurkat cell line (ab271143). 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, 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
Lck initiates and propagates T-cell receptor (TCR) signaling cascades. It interacts with the CD4 and CD8 co-receptors and triggers the activation of other kinases within the cell. Lck as part of a signaling complex associates with proteins such as ZAP-70 and LAT facilitating the assembly of larger molecular machines necessary for immune responses. Its activity is tightly regulated by phosphorylation with key sites being tyrosine 394 and 505.
Pathways
The Lck protein plays an essential role in TCR signaling and immune responses. It actively participates in the Lck signaling pathway setting off cascades that lead to T-cell activation and differentiation. It interacts closely with other proteins like Fyn and Src making sure that the pathway progresses correctly. In conjunction with the immune synapse Lck ensures the transmission of signals that allow the T-cells to respond to external antigens efficiently.
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
Complementary Products
![Western blot - Anti-Lck antibody [EPR20798-107] (AB227975)](https://content.abcam.com/products/images/lck-antibody-epr20798-107-ab227975--western-blot-img19738.jpg)
Product promise
Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.
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