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AB273809

Human LCK knockout Jurkat cell lysate

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LCK KO cell lysate available now. KO validated by Next Generation Sequencing, Western blot. Free of charge wild type control included. 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

3 Images
Western blot - Human LCK knockout Jurkat cell lysate (AB273809)
  • WB

Lab

Western blot - Human LCK knockout Jurkat cell lysate (AB273809)

Lane 1 : Wild-type Jurkat cell lysate 20 μg
Lane 2 : Lck knockout Jurkat cell lysate 20 μg
Lane 3 : Ramos cell lysate 20 μg
Lane 4 : A549 cell lysate 20 μg
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 °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-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 (<a href='/en-us/products/cell-lines/human-lck-knockout-jurkat-cell-line-ab273855'>ab273855</a>)

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

Western blot - Human LCK knockout Jurkat cell lysate (AB273809)
  • WB

Lab

Western blot - Human LCK knockout Jurkat cell lysate (AB273809)

Lane 1 : Wild-type Jurkat cell lysate 20 μg
Lane 2 : Lck knockout Jurkat cell lysate 20 μg
Lane 3 : Ramos cell lysate 20 μg
Lane 4 : A549 cell lysate 20 μg
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 °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-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 (<a href='/en-us/products/cell-lines/human-lck-knockout-jurkat-cell-line-ab273855'>ab273855</a>)

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

Next Generation Sequencing - Human LCK knockout Jurkat cell lysate (AB273809)
  • NGS

Supplier Data

Next Generation Sequencing - Human LCK knockout Jurkat cell lysate (AB273809)

Knockout achieved by CRISPR/Cas9; X = 1 bp insertion, 2 bp insertion; Frameshift : 99%

Key facts

Cell type

Jurkat

Species or organism

Human

Tissue

Blood

Knockout validation

Next Generation Sequencing,Western blot

Mutation description

Knockout achieved by CRISPR/Cas9; X = 1 bp insertion, 2 bp insertion; Frameshift: 99%

Disease

Non-Hodgkin Lymphoma

Reactivity data

{ "title": "Reactivity Data", "filters": { "stats": ["", "Reactivity", "Dilution Info", "Notes"] }, "values": { "WB": { "reactivity":"TESTED_AND_REACTS", "dilution-info":"", "notes":"<p></p>" } } }

Product details

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?

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Properties and storage information

Gene name
LCK
Gene editing type
Knockout
Gene editing method
CRISPR technology
Knockout validation
Next Generation Sequencing, Western blot
Shipped at conditions
Ambient - Can Ship with Ice
Appropriate short-term storage conditions
-20°C
Appropriate long-term storage conditions
-20°C

Supplementary information

This supplementary information is collated from multiple sources and compiled automatically.

Lck also known as lymphocyte-specific protein tyrosine kinase plays an important role in T-cell activation. It serves as an enzyme and is integral in the signal transduction of the immune system. Lck has a molecular weight of approximately 56 kDa. It is expressed predominantly in T lymphocytes and is part of the Src family kinases. This protein is important in phosphorylating substrates leading to downstream signaling events that are necessary for the proper immune function.
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.

Defective Lck activity can contribute to immunodeficiencies and autoimmune diseases. Proper Lck function is linked to conditions like Severe Combined Immunodeficiency (SCID) and lupus. Dysregulation in Lck activity may also affect proteins such as ZAP-70 worsening disease progression. Understanding Lck's participation in these conditions can lead to better therapeutic interventions and aid in developing targeted treatments for immune-related disorders.

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 protocols

Product promise

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