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Products:Immunology >> Adaptive Immunity >> T Cells >> CD
Overview
Product name
Anti-Lck antibody
See all Lck products (18) ...
Description
Rabbit polyclonal to Lck
Specificity
This antibody was designed to recognise the activated form of Lck and was raised for that purpose against the linker that connects the SH2 domain and the tyrosine kinase domain of the kinase. This linker sits in the SH3 substrate binding site in the inactive kinase. The position of the linker is based on the crystal structure of inactive Src. However, the antibody does not seem to distinguish the active and inactive forms of Lck. In IF, it recognises kinase dead Lck at the same concentration as constitutively active Lck. In western blot, the antibody is highly specific for Lck versus other tyrosine kinases (see blot). Even, if the antibody was specific for active Lck in IF , it would be expected to recognise both active and inactive forms in Western blot. This is because the linker that the antibody recognises is present in both active and inactive Lck, it was just expected to be concealed in native form inactive Lck.
Tested applications
IHC-P, WB, IFmore details
Cross reactivity
Reacts with
Mouse, African Green Monkey
Predicted to work with
Chicken, Human
Immunogen
Synthetic peptide conjugated to KLH derived from within residues 200 - 300 of Human Lck.
(Peptide available as ab13750.)
Properties
Form
Liquid
Storage instructions
Store at +4°C short term (1-2 weeks). Aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.
Storage buffer
Preservative: 0.02% Sodium Azide
Constituents: 1% BSA, PBS, pH 7.4
Concentration
Concentration information loading...
Purity
Immunogen affinity purified
Primary antibody notes
Lck (p56lck), a member of the Src family of non-receptor tyrosine protein kinases, is expressed predominantly in T cells.
Clonality
Polyclonal
Isotype
IgG
Research areas
Immunology >> Secreted Molecules >> Other secreted molecules
Signal Transduction >> Protein Phosphorylation >> Tyrosine Kinases >> Src Family
Immunology >> Immunoglobulins >> Receptors
Immunology >> Adaptive Immunity >> T Cells >> CD
Applications
Show applications key
Our Abpromise guarantee covers the use of ab3885 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
- 2 Images
WB
IF
Application notes
IF: 1/200 - 1/500.
IHC-P: 1/2000 (See abreview).
WB: 1/1000. Predicted molecular weight: 57 kDa. Can be blocked with Lck peptide (ab13750) or Lck peptide (ab13750).
Not yet tested in other applications.
Optimal dilutions/concentrations should be determined by the end user.
Target
Function
Tyrosine kinase that plays an essential role for the selection and maturation of developing T-cell in the thymus and in mature T-cell function. Is constitutively associated with the cytoplasmic portions of the CD4 and CD8 surface receptors and plays a key role in T-cell antigen receptor(TCR)-linked signal transduction pathways. 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, and thereby recruits the associated LCK to the vicinity of the TCR/CD3 complex. LCK then phosphorylates tyrosines residues within the immunoreceptor tyrosines-based activation motifs (ITAMs) in the cytoplasmic tails of the TCRgamma chains and CD3 subunits, initiating the TCR/CD3 signaling pathway. In addition, contributes to signaling by other receptor molecules. Associates directly with the cytoplasmic tail of CD2, and upon engagement of the CD2 molecule, LCK undergoes hyperphosphorylation and activation. Also plays a role in the IL2 receptor-linked signaling pathway that controls 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 RUNX3.
Tissue specificity
Expressed specifically in lymphoid cells.
Involvement in disease
Note=A chromosomal aberration involving LCK is found in leukemias. Translocation t(1;7)(p34;q34) with TCRB.
Sequence similarities
Belongs to the protein kinase superfamily. Tyr protein kinase family. SRC subfamily.
Contains 1 protein kinase domain.
Contains 1 SH2 domain.
Contains 1 SH3 domain.
Domain
The SH2 domain mediates interaction with SQSTM1. Interaction is regulated by Ser-59 phosphorylation.
Post-translational
modifications
Phosphorylated on Tyr-394, which increases enzymatic activity (By similarity). Phosphorylated on Tyr-505, which decreases activity.
Cellular localization
Cytoplasm. Cell membrane. Present in lipid rafts in an unactive form.
Target information above from: UniProt accessionP06239
The UniProt Consortium
The Universal Protein Resource (UniProt) in 2010
Nucleic Acids Res. 38:D142-D148 (2010).
Alternative names
- LCK antibody
- Lck p56 antibody
- LCK_HUMAN antibody
see all
Database links
- Entrez Gene: 3932 Human
- Entrez Gene: 16818 Mouse
- GenBank: NM_005356 Human
- GenBank: NM_010693 Mouse
- Omim: 153390 Human
- SwissProt: P42683 Chicken
- SwissProt: P06239 Human
- SwissProt: P06240 Mouse
see all
Anti-Lck antibody images:
Western blot - Lck antibody (ab3885)
All lanes : Anti-Lck antibody (ab3885) at 1/1000 dilution
Lane 1 : COS cells transfected with mouse Fyn
Lane 2 : COS cells transfected with mouse Lck
Predicted band size : 57 kDa
Immunocytochemistry/ Immunofluorescence - Anti-Lck antibody (ab3885)
Confocal immunofluorescence image of a COS cell transfected with a mouse Lck construct and stained with ab3885 at 1/200.
In untransfected cells, some nuclear staining is also visible as well as Lck staining.
Western blot - Lck antibody (ab3885)
All lanes : Anti-Lck antibody (ab3885) at 0.87 µg/ml
Lane 1 : HeLa nuclear extract
Lane 2 : HeLa whole cell extract
Lane 3 : A431 cell extract
Lane 4 : Jurkat cell lysate
Lane 5 : HEK293 cell lysate
Lane 6 : HeLa nuclear extract with Lck peptide (ab13750) at 1 µg/ml
Lane 7 : HeLa whole cell extract with Lck peptide (ab13750) at 1 µg/ml
Lane 8 : A431 cell extract with Lck peptide (ab13750) at 1 µg/ml
Lane 9 : Jurkat cell lysate with Lck peptide (ab13750) at 1 µg/ml
Lane 10 : HEK293 cell lysate with Lck peptide (ab13750) at 1 µg/ml
Predicted band size : 57 kDa
Observed band size : 57 kDa
Additional bands at : 25 kDa (possible non-specific binding),35 kDa (possible non-specific binding).
ab3885 recognises a band at approximately 57 kDa, corresponding to Lck in HeLa nuclear (lane1), HeLa whole cell (lane2) and Jurkat cell (lane4) lysates. The recognition of these bands is succesfully blocked using the immunizing peptide (lanes 6, 7, 9 respectively). Lck is not detected using ab3885 in A431 (lane3) or HEK293 (lane5) cells under these conditions.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Lck antibody (ab3885)
ab3885 staining Lck in human thymus tissue section by Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections). Tissue underwent fixation in formaldehyde, heat mediated antigen retrieval and blocking was done for 15 minutes at 20°C (5 minutes of peroxidase block then 10 minutes of protein block) . The primary antibody was diluted, 1/2000 and incubated with sample for 45 minutes at 20°C. A HRP conjugated goat polyclonal to rabbit IgG was used, undiluted as secondary.
This image is courtesy of an Abreview submitted by Antibody Solutions Ltd.
References for Anti-Lck antibody (ab3885)
This product has been referenced in:
- Rüder Cet al. The tumor-associated antigen EBAG9 negatively regulates the cytolytic capacity of mouse CD8+ T cells. J Clin Invest 119:2184-203 (2009). IHC (PFA fixed); Mouse.Read more (PubMed: 19620783) »
See 1 publication for this product
Publishing research using ab3885? Please let us know so that we can cite the reference in this datasheet
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"