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Alexa Fluor® 488 anti-Lck Phospho (Tyr394) Antibody

Pricing & Availability
Clone
A18002D (See other available formats)
Regulatory Status
RUO
Other Names
Lsk, p56lck, LCK Proto-Oncogene, Src Family Tyrosine Kinase, Tyrosine-protein kinase LCK, Leukocyte C-terminal Src kinase, LSK, Lymphocyte cell-specific protein-tyrosine kinase, T cell-specific protein-tyrosine kinase
Isotype
Mouse IgG1, κ
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Product Citations
publications
A. A18002D_A488_LckPhosphoTyr394_ICFC_062525
Jurkat cells serum-starved overnight and then treated with 0.03% H2O2 for 5 minutes to activate Lck (positive cell line, filled histogram) or left untreated (negative cell line, open histogram) were fixed and permeabilized using Cyto-Fast™ Fix/Perm Buffer Set (Cat. No. 426803) and intracellularly stained with Alexa Fluor® 488 Lck Phospho (Tyr394) or Alexa Fluor® 488 Mouse IgG1, κ Isotype Control (open histogram, dashed line, representative histogram for both untreated and treated cells) (Cat. No. 400134).
  • A. A18002D_A488_LckPhosphoTyr394_ICFC_062525
    Jurkat cells serum-starved overnight and then treated with 0.03% H2O2 for 5 minutes to activate Lck (positive cell line, filled histogram) or left untreated (negative cell line, open histogram) were fixed and permeabilized using Cyto-Fast™ Fix/Perm Buffer Set (Cat. No. 426803) and intracellularly stained with Alexa Fluor® 488 Lck Phospho (Tyr394) or Alexa Fluor® 488 Mouse IgG1, κ Isotype Control (open histogram, dashed line, representative histogram for both untreated and treated cells) (Cat. No. 400134).
  • B. A18002D_A488_LckPhosphoTyr394_ICC_062525
    Jurkat cells were serum starved for 24 hours then treated for 5 minutes with 0.03% H2O2 to activate Lck (panel B) or left untreated (panel A). Cells were fixed with 4% PFA Fixation Buffer (Cat. No. 420801) for 10 minutes and permeabilized with 0.5% Triton X-100 for 10 minutes and blocked with 5% FBS for 1 hour at room temperature. Cells were then stained with Alexa Fluor® 488 Lck Phospho (Tyr394) (clone A18002D). Nuclei were counterstained with DAPI (Cat. No. 422801). The images were captured on a Revvity Operetta CLS™ High Content Analysis System with a 63X objective and merged. Scale bar: 50 µm
Compare all formats See Alexa Fluor® 488 spectral data
Cat # Size Price Save
933107 25 µg ¥35,200
933108 100 µg ¥79,200
Description

The Src family tyrosine kinase p56Lck (Lck) is a non-receptor tyrosine kinase that plays a critical role in T cell selection and maturation within the thymus, and also in the function of mature T cells. Lck, which is constitutively bound to cytosolic domains of CD4 and CD8 surface receptors, plays an essential role in T cell receptor (TCR) signaling. Engagement of the TCR with peptide antigen-loaded MHC complexes results in the recruitment of CD4- and CD8-bound Lck to the TCR/CD3 signaling complex. Lck then transphosphorylates TCR-gamma chains and CD3 subunits, thereby activating the TCR/CD3 signaling pathway and leading to the recruitment and subsequent phosphorylation of Zap70 by Lck. Lck also plays an important role in interleukin-2 signaling that regulates the T cell proliferative response. Phosphorylation of Lck by CSK at tyrosine 505 negatively regulates the kinase, and is proposed to generate a closed, inactive conformation of the protein. Conversely, phosphorylation at tyrosine 394 in the activation loop of Lck greatly stimulates enzymatic activity. In many types of cancer, Lck is a proliferative and anti-apoptotic driver, and has been proposed as a target for therapeutic intervention.

Product Details
Technical data sheet

Product Details

Verified Reactivity
Human, Mouse
Antibody Type
Monoclonal
Host Species
Mouse
Immunogen
Synthetic peptide corresponding to human Src phosphorylated at tyrosine 419
Formulation
Phosphate-buffered solution, pH 7.2, containing 0.09% sodium azide
Preparation
The antibody was purified by affinity chromatography and conjugated with Alexa Fluor® 488 under optimal conditions.
Concentration
0.5 mg/mL
Storage & Handling
The antibody solution should be stored undiluted between 2°C and 8°C, and protected from prolonged exposure to light. Do not freeze.
Application

ICFC - Quality tested
ICC - Verified
Recommended Usage

Each lot of this antibody is quality control tested by intracellular immunofluorescent staining with flow cytometric analysis. For flow cytometric staining, the suggested use of this reagent is ≤ 0.5 µg per million cells in 100 µL volume. For immunocytochemistry, a concentration range of 1.0 - 10.0 μg/mL is recommended. It is recommended that the reagent be titrated for optimal performance for each application.

* Alexa Fluor® 488 has a maximum emission of 519 nm when it is excited at 488 nm.


Alexa Fluor® and Pacific Blue™ are trademarks of Life Technologies Corporation.

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Excitation Laser
Blue Laser (488 nm)
Application Notes

This clone may recognize other Src family members. This clone was developed using a synthetic peptide corresponding to human Src phosphorylated at tyrosine 419, which displays high sequence homology with the region surrounding Lck tyrosine 394. To confirm specificity to Lck Phospho (Tyr394), the cell line J.Cam1.6 was used. This cell line is a clonal derivative of Jurkat cells that harbors a truncated LCK allele with defective protein expression (Straus, et al. 1992. Cell. 70:585).

Additional Product Notes

For use in immunocytochemistry (ICC), it is recommended to fix/permeabilize with either of the following:
-Fixation buffer (Cat. No. 420801) with 0.5 % Triton X-100
-Fixation buffer (Cat. No. 420801) with 100% ice-cold methanol
-100% ice-cold methanol only

Antigen Details

Structure
Lck is a 509 amino acid protein with a predicted molecular weight of 58 kD
Distribution

Plasma-membrane associated/T cells

Function
Protein Tyrosine Kinase
Interaction
CD4, CD8, CD2, ZAP-70, LAT, CD45, CSK, SHP1, FYN
Cell Type
T cells
Biology Area
Cell Biology, Signal Transduction
Molecular Family
Protein Kinases/Phosphatase
Antigen References
  1. Phillipsen L, et al. 2017. Sci Signal. 10:eaaf4736.
  2. Moogk D, et al. 2016. J Immunol. 197:644.
  3. Klammt C, et al. 2015. Nat Immunol. 16:961.
  4. Cancer Genome Atlas Network. 2015. Cell. 161:1681.
  5. Casas J, et al. 2014. Nat Commun. 5:5624
  6. McNeill L, et al. 2007. Immunnity. 27:425.
  7. Lefebvre DC, et al. 2003. Biochim Biophys Acta. 1650:40.
  8. Kabouridis PS. et al. 2003. Biochem J. 371:907.
  9. Abraham N, et al. 1990. Mol. Cell. Biol. 10:5197.
Gene ID
3932 View all products for this Gene ID
UniProt
View information about Lck Phospho Tyr394 on UniProt.org

Related FAQs

There are no FAQs for this product.
Go To Top Version: 1    Revision Date: 06/25/2025

For Research Use Only. Not for diagnostic or therapeutic use.

 

This product is supplied subject to the terms and conditions, including the limited license, located at www.biolegend.com/terms) ("Terms") and may be used only as provided in the Terms. Without limiting the foregoing, BioLegend products may not be used for any Commercial Purpose as defined in the Terms, resold in any form, used in manufacturing, or reverse engineered, sequenced, or otherwise studied or used to learn its design or composition without express written approval of BioLegend. Regardless of the information given in this document, user is solely responsible for determining any license requirements necessary for user’s intended use and assumes all risk and liability arising from use of the product. BioLegend is not responsible for patent infringement or any other risks or liabilities whatsoever resulting from the use of its products.

 

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This data display is provided for general comparisons between formats.
Your actual data may vary due to variations in samples, target cells, instruments and their settings, staining conditions, and other factors.
If you need assistance with selecting the best format contact our expert technical support team.

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