AB273855

Human LCK knockout Jurkat cell line

Advanced Validation
詳細を見る

Be the first to review this product! Submit a review

(0 Publication)

LCK KO cell line available to order. KO validated by Next Generation Sequencing, Western blot. Free of charge wild type control available. Knockout achieved by CRISPR/Cas9; X = 1 bp insertion, 2 bp insertion; Frameshift: 99%. To order both knockout and wild-type control cells: select '2 x 1000000 Cells/vial'. To order only knockout cells: select '1000000 Cells/vial'.

別名を表示する

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

4 Images

Lab

Western blot - Human LCK knockout Jurkat cell line (AB273855)

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='/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:

Western blot - Human LCK knockout Jurkat cell lysate (<a href='/products/cell-lysates/human-lck-knockout-jurkat-cell-lysate-ab273809'>ab273809</a>) at 20 µg

Lane 3:

Ramos cell lysate at 20 µg

Lane 4:

A549 cell lysate at 20 µg

Secondary

Lanes 1 - 4:

Western blot - Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-irdye-800cw-preadsorbed-ab216773'>ab216773</a>) at 1/20000 dilution

Lanes 1 - 4:

Western blot - Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (<a href='/products/secondary-antibodies/goat-mouse-igg-h-l-irdye-680rd-preadsorbed-ab216776'>ab216776</a>) at 1/20000 dilution

Predicted band size: 58 kDa

Observed band size: 60 kDa,37 kDa

false

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='/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

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

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%

Key facts

細胞タイプ

Jurkat

生物種

Human

組織

Blood

製品の状態

Liquid

form

ノックアウト検証方法

Next Generation Sequencing,Western blot

ノックアウト変異

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

疾病

Non-Hodgkin Lymphoma

style

left-column

Reactivity data

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

style

left-column

製品の詳細

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.

style

left-column

製品内容

{ "values": { "2x1000000Cellsvial": { "sellingSize": "2 x 1000000 Cells/vial", "publicAssetCode":"ab273855-2x1000000Cells_vial", "assetComponentDetails": [ { "size":"1 x 1000000 Cells/vial", "name":"ab273855 Human LCK knockout Jurkat cell line", "number":"AB273855-CMP01" }, { "size":"1 x 1000000 Cells/vial", "name":"ab271143 Human wild-type Jurkat cell line", "number":"AB273855-CMP02" } ] }, "1000000Cellsvial": { "sellingSize": "1000000 Cells/vial", "publicAssetCode":"ab273855-1000000Cells_vial", "assetComponentDetails": [ { "size":"1 x 1000000 Cells/vial", "name":"ab273855 Human LCK knockout Jurkat cell line", "number":"AB273855-CMP01", "productcode":"" } ] } } }

style

left-column

出荷温度及び保存条件

遺伝子名

LCK

遺伝子編集のタイプ

Knockout

遺伝子編集の方法

CRISPR technology

ノックアウト検証方法

Next Generation Sequencing, Western blot

出荷温度

Dry Ice

短期保存温度

-196°C

長期保存温度

-196°C

style

left-column

取り扱い方法

初回取り扱いガイドライン

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.

継代培養ガイドライン

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.

培養培地

RPMI + 10% FBS

凍結保存培地

Cell Freezing Medium-DMSO Serum free media, contains 8.7% DMSO in MEM supplemented with methyl cellulose.

style

left-column

補足情報

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.

style

left-column

style

left-column

Quality control

STR analysis

CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX

style

left-column

Cell culture

Biosafety level

EU: 1 US: 1

Adherent/suspension

Suspension

Gender

Male

style

left-column

製品プロトコール

Visit the General protocols
Visit the Troubleshooting

style

left-column

ターゲットの情報

See full target information LCK

style

left-column

style

left-column

style

right-column

Abcam product promise

当社は、高品質な試薬を通じてお客様の研究を力強くサポートすることをお約束いたします。ご使用いただく各段階で、常にお客様をサポートできる体制を整えております。万が一、製品が期待通りに機能しない場合は、「Abcam Product Promise」による当社保証制度に基づき、安心してご利用いただけます。

保証に関する詳細については利用規約をご確認ください。

Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.

For licensing inquiries, please contact partnerships@abcam.com

Human LCK knockout Jurkat cell line