Skip to main content

Phospho-Lck (Y394) ELISA Kit is a Semi-quantitative ELISA kit for the measurement of Phospho-Lck (Y394) in Human in Cell Lysate samples.

Be the first to review this product! Submit a review

Images

Sandwich ELISA - Phospho-Lck (Y394) ELISA Kit (AB279848), expandable thumbnail
  • Sandwich ELISA - Phospho-Lck (Y394) ELISA Kit (AB279848), expandable thumbnail
  • Sandwich ELISA - Phospho-Lck (Y394) ELISA Kit (AB279848), expandable thumbnail

Key facts

Detection method
Colorimetric
Sample types
Cell Lysate
Assay type
Semi-quantitative
Reactive species
Human

Reactivity data

Application
ELISA
Reactivity
Reacts
Dilution info
-
Notes

-

Target data

Function

Non-receptor tyrosine-protein kinase that plays an essential role in the selection and maturation of developing T-cells in the thymus and in the function of mature T-cells. Plays a key role in T-cell antigen receptor (TCR)-linked signal transduction pathways. Constitutively associated with the cytoplasmic portions of the CD4 and CD8 surface receptors. Association of the TCR with a peptide antigen-bound MHC complex facilitates the interaction of CD4 and CD8 with MHC class II and class I molecules, respectively, thereby recruiting the associated LCK protein to the vicinity of the TCR/CD3 complex. LCK then phosphorylates tyrosine residues within the immunoreceptor tyrosine-based activation motifs (ITAM) of the cytoplasmic tails of the TCR-gamma chains and CD3 subunits, initiating the TCR/CD3 signaling pathway. Once stimulated, the TCR recruits the tyrosine kinase ZAP70, that becomes phosphorylated and activated by LCK. Following this, a large number of signaling molecules are recruited, ultimately leading to lymphokine production. LCK also contributes to signaling by other receptor molecules. Associates directly with the cytoplasmic tail of CD2, which leads to hyperphosphorylation and activation of LCK. Also plays a role in the IL2 receptor-linked signaling pathway that controls the T-cell proliferative response. Binding of IL2 to its receptor results in increased activity of LCK. Is expressed at all stages of thymocyte development and is required for the regulation of maturation events that are governed by both pre-TCR and mature alpha beta TCR. Phosphorylates other substrates including RUNX3, PTK2B/PYK2, the microtubule-associated protein MAPT, RHOH or TYROBP. Interacts with FYB2 (PubMed:27335501).

Alternative names

What's included?

1 x 96 Tests
Components
100X HRP-Conjugated Streptavidin Concentrate
1 x 200 µL
20X Wash Buffer
1 x 25 mL
2X Cell lysate buffer
1 x 5 mL
5X Assay Diluent
1 x 15 mL
Biotinylated anti-phospho-Lck (Y394)-antibody
2 x 1 Vial
Pan-Lck Coated Microplate
1 x 1 Unit
Positive Control - Jurkat cell lysate
1 x 1 Vial
Stop Solution
1 x 8 mL
TMB One-Step Substrate Reagent
1 x 12 mL

Recommended products

Phospho-Lck (Y394) ELISA Kit is a Semi-quantitative ELISA kit for the measurement of Phospho-Lck (Y394) in Human in Cell Lysate samples.

Key facts

Detection method
Colorimetric
Sample types
Cell Lysate
Assay type
Semi-quantitative
Reactive species
Human
Assay Platform
Pre-coated microplate (12 x 8 well strips)

Storage

Shipped at conditions
Blue Ice
Appropriate short-term storage conditions
-20°C
Appropriate long-term storage conditions
-20°C
Storage information
-20°C

Notes

Phospho-Lck (Y394) ELISA Kit (ab279848) is a very rapid, convenient, and sensitive assay kit that can monitor the activation or function of important biological pathways in human cell lysates. By determining phosphorylated Lck protein in your experimental model system, you can verify pathway activation in your cell lysates. You can simultaneously measure numerous different cell lysates without spending excess time and effort in performing a Western Blotting analysis.

This Sandwich ELISA kit is an in vitro enzyme-linked immunosorbent assay for the measurement of human phospho-Lck. An anti-pan Lck antibody has been coated onto a 96-well plate. Samples are pipetted into the wells and Lck present in a sample is bound to the wells by the immobilized antibody. The wells are washed and biotinylated anti-phospho-Lck (Y394) antibody is used to detect phosphorylated Lck. After washing away unbound antibody, HRP-conjugated Streptavidin is pipetted into the wells. The wells are again washed, a TMB substrate solution is added to the wells and color develops in proportion to the amount of Lck (Y394) bound. The Stop Solution changes the color from blue to yellow, and the intensity of the color is measured at 450 nm.

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.

Supplementary info

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

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.

Associated diseases and disorders

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.

Product promise

We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.

In the unlikely event of one of our products not working as expected, you are covered by our product promise.

Full details and terms and conditions can be found here:
Terms & Conditions.

3 product images

  • Sandwich ELISA - Phospho-Lck (Y394) ELISA Kit (ab279848), expandable thumbnail

    Sandwich ELISA - Phospho-Lck (Y394) ELISA Kit (ab279848)

    Positive Control.

    Jurkat cells were treated with Pervanadate at 37°C for 10 mins.

    Solubilize cells at 4 x 107 cells/ml in Cell Lysate Buffer.

    Serial dilutions of lysates were analyzed in this ELISA.

  • Sandwich ELISA - Phospho-Lck (Y394) ELISA Kit (ab279848), expandable thumbnail

    Sandwich ELISA - Phospho-Lck (Y394) ELISA Kit (ab279848)

    Jurkat cells treated with/without Pervanadate. Jurkat cells were untreated or treated with Pervanadate for 10 minutes.

  • Sandwich ELISA - Phospho-Lck (Y394) ELISA Kit (ab279848), expandable thumbnail

    Sandwich ELISA - Phospho-Lck (Y394) ELISA Kit (ab279848)

    Jurkat cells treated with/without Pervanadate. Jurkat cells were untreated or treated with Pervanadate for 10 minutes.

Downloads

Product protocols

For this product, it's our understanding that no specific protocols are required. You can:

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