Anti-Lck antibody [EPR20798-107] (ab227975)

Rabbit Recombinant Monoclonal LCK antibody. Suitable for IP, WB, ICC/IF, Flow Cyt (Intra), IHC-P and reacts with Human, Mouse, Rat samples. Cited in 1 publication.

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Images

Immunoprecipitation - Anti-Lck antibody [EPR20798-107] (AB227975), expandable thumbnail

Publications

Key facts

Isotype: IgG
Host species: Rabbit
Storage buffer: pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Form: Liquid
Clonality: Monoclonal

Immunogen

The exact immunogen used to generate this antibody is proprietary information.

Reactivity data

Select an application

Product promiseTestedExpectedPredictedNot recommended

	IP	WB	ICC/IF	Flow Cyt (Intra)	IHC-P
Human	Tested	Tested	Tested	Tested	Tested
Mouse	Expected	Tested	Tested	Tested	Tested
Rat	Expected	Tested	Expected	Expected	Tested

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info 1/30	Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Expected
Expected

Species	Dilution info	Notes
Species Mouse, Rat	Dilution info Use at an assay dependent concentration.	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Mouse	Dilution info 1/1000	Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Species Rat	Dilution info 1/1000	Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Species Human	Dilution info 1/1000	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Mouse	Dilution info 1/100	Notes -
Species Human	Dilution info 1/100	Notes -

Expected
Expected

Species	Dilution info	Notes
Species Rat	Dilution info Use at an assay dependent concentration.	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Mouse	Dilution info 1/500	Notes -
Species Human	Dilution info 1/500	Notes -

Expected
Expected

Species	Dilution info	Notes
Species Rat	Dilution info Use at an assay dependent concentration.	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Mouse	Dilution info 1/1000	Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Species Rat	Dilution info 1/1000	Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Species Human	Dilution info 1/1000	Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Associated Products

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Target data

See full target information LCK

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

Tyrosine-protein kinase Lck, Leukocyte C-terminal Src kinase, Lymphocyte cell-specific protein-tyrosine kinase, Protein YT16, Proto-oncogene Lck, T cell-specific protein-tyrosine kinase, p56-LCK, LSK, LCK

Recommended products

Rabbit Recombinant Monoclonal LCK antibody. Suitable for IP, WB, ICC/IF, Flow Cyt (Intra), IHC-P and reacts with Human, Mouse, Rat samples. Cited in 1 publication.

Key facts

Isotype: IgG
Host species: Rabbit
Storage buffer: pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Form: Liquid
Clonality: Monoclonal
Immunogen: The exact immunogen used to generate this antibody is proprietary information.
Clone number: EPR20798-107
Purification technique: Affinity purification Protein A
Concentration

Storage

Shipped at conditions: Blue Ice
Appropriate short-term storage duration: 1-2 weeks
Appropriate short-term storage conditions: +4°C
Appropriate long-term storage conditions: -20°C
Aliquoting information: Upon delivery aliquot
Storage information: Avoid freeze / thaw cycle

Notes

Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

This product is a recombinant monoclonal antibody, which offers several advantages including:

- High batch-to-batch consistency and reproducibility
- Improved sensitivity and specificity
- Long-term security of supply
- Animal-free batch production

For more information, read more on recombinant antibodies.

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

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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:
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12 product images

Immunoprecipitation - Anti-Lck antibody [EPR20798-107] (ab227975)
Lck was immunoprecipitated from 0.35 mg of Jurkat (human T cell leukemia cell line from peripheral blood) whole cell lysate with ab227975 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab227975 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (VeriBlot for IP Detection Reagent (HRP) ab131366), was used for detection at 1/10000 dilution.
Lane 1: Jurkat whole cell lysate 10 μg (Input).
Lane 2: ab227975 IP in Jurkat whole cell lysate.
Lane 3: Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of ab227975 in Jurkat whole cell lysate.
Exposure time: 8 seconds.
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
All lanes: Immunoprecipitation - Anti-Lck antibody [EPR20798-107] (ab227975)
Developed using the ECL technique.
Predicted band size: 58 kDa
Observed band size: 58 kDa
Western blot - Anti-Lck antibody [EPR20798-107] (ab227975)
False colour image of Western blot: Anti-Lck antibody [EPR20798-107] staining at 1/1000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (Anti-GAPDH antibody [6C5] - Loading Control 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 Human LCK knockout Jurkat cell line ab273855 (knockout cell lysate Human LCK knockout Jurkat 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 (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776) at 1/20000 dilution.
All lanes: Western blot - Anti-Lck antibody [EPR20798-107] (ab227975) 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 (Human LCK knockout Jurkat cell line ab273855)
Lane 3: Ramos cell lysate at 20 µg
Lane 4: A549 cell lysate at 20 µg
Performed under reducing conditions.
Predicted band size: 58 kDa
Observed band size: 60 kDa
Western blot - Anti-Lck antibody [EPR20798-107] (ab227975)
Exposure times: Lanes 1,2,7: 5 seconds; Lanes 3,4,8: 1 second; Lane 5: 3 seconds; Lane 6: 10 seconds.
Blocking/Dilution buffer: 5% NFDM/TBST.
All lanes: Western blot - Anti-Lck antibody [EPR20798-107] (ab227975) at 1/1000 dilution
Lane 1: Human thymus tissue lysate at 20 µg
Lane 2: Human tonsil tissue lysate at 20 µg
Lane 3: WEHI-231 (mouse lymphoblast B cell lymphoma cell line) whole cell lysate at 20 µg
Lane 4: Rat thymus tissue lysate at 20 µg
Lane 5: Mouse lymph node tissue lysate at 20 µg
Lane 6: Raji (human Burkitt's lymphoma cell line) whole cell lysate at 20 µg
Lane 7: Ramos (human Burkitt's lymphoma cell line) whole cell lysate at 20 µg
Lane 8: Jurkat (human T cell leukemia cell line from peripheral blood) whole cell lysate at 20 µg
Secondary
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution
Developed using the ECL technique.
Predicted band size: 58 kDa
Observed band size: 58 kDa
Flow Cytometry (Intracellular) - Anti-Lck antibody [EPR20798-107] (ab227975)
Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed, 90% methanol-permeabilized WEHI-231 (mouse lymphoblast B cell lymphoma cell line) cell line labeling Lck with ab227975 at 1/500 dilution (red) compared with a Rabbit IgG, monoclonal [EPR25A] - Isotype Control (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) (black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (blue). Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) at 1/2000 dilution was used as the secondary antibody.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Lck antibody [EPR20798-107] (ab227975)
Immunohistochemical analysis of paraffin-embedded rat colon tissue labeling Lck with ab227975 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP), ready to use. Membranous and cytoplasmic staining in T cells of rat colon is observed (PMID: 16769579). Counter stained with Hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP), ready to use.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Flow Cytometry (Intracellular) - Anti-Lck antibody [EPR20798-107] (ab227975)
Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed, 90% methanol permeabilized Jurkat (human T cell leukemia cell line from peripheral blood) cell line labeling Lck with ab227975 at 1/500 dilution (red) compared with a Rabbit IgG, monoclonal [EPR25A] - Isotype Control (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) (black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (blue). Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) at 1/2000 dilution was used as the secondary antibody.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Lck antibody [EPR20798-107] (ab227975)
Immunohistochemical analysis of paraffin-embedded mouse spleen tissue labeling Lck with ab227975 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP), ready to use. Membranous and cytoplasmic staining in mouse spleen reactive lymph node and T cells is observed (PMID: 16769579). Counter stained with Hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP), ready to use.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Lck antibody [EPR20798-107] (ab227975)
Immunohistochemical analysis of paraffin-embedded human diffuse large B-cell lymphoma tissue labeling Lck with ab227975 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP), ready to use. Membranous and cytoplasmic staining in human diffuse large B-cell lymphoma is observed (PMID:16769579). Counter stained with Hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP), ready to use.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Immunocytochemistry/ Immunofluorescence - Anti-Lck antibody [EPR20798-107] (ab227975)
Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized WEHI-231 (mouse lymphoblast B cell lymphoma cell line) cells labeling Lck with ab227975 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing membranous staining on WEHI-231 cell line.
The nuclear counter stain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) (Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker ab195889) at 1/200 dilution (red).
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) at 1/1000 dilution.
Immunocytochemistry/ Immunofluorescence - Anti-Lck antibody [EPR20798-107] (ab227975)
Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized Ramos (human Burkitt's lymphoma cell line) cells labeling Lck with ab227975 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing membranous staining on Ramos cell line.
The nuclear counter stain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) (Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker ab195889) at 1/200 dilution (red).
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) at 1/1000 dilution.
Western blot - Anti-Lck antibody [EPR20798-107] (ab227975)
In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (Anti-GAPDH antibody [EPR16891] - Loading Control ab181602) staining at 1/200000 dilution.
In Western blot, Anti-Lck antibody - Total protein control (ab227975) staining at 1/1000 dilution.
All lanes: Western blot - Anti-Lck (phospho Y394) antibody [EPR27974-93] (Anti-Lck (phospho Y394) antibody [EPR27974-93] ab318960) at 1/1000 dilution
Lane 1: COLO 205 (human colon adenocarcinoma tumor ascitic epithelial cell) whole cell lysate (untreated membrane) at 20 µg with NFDM/TBST
Lane 2: COLO 205 whole cell lysate (alkaline phosphatase treated membrane) at 20 µg with NFDM/TBST
Secondary
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution
Observed band size: 60 kDa, 36 kDa
Exposure time: 180s
Western blot - Anti-Lck antibody [EPR20798-107] (ab227975)
The expression of Lck (phospho Y394) is downregulated in response to Dasatinib treatment.
In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (Anti-GAPDH antibody [EPR16891] - Loading Control ab181602) staining at 1/200000 dilution.
In Western blot, Anti-Lck antibody - Total protein control (ab227975) staining at 1/1000 dilution.
All lanes: Western blot - Anti-Lck (phospho Y394) antibody [EPR27974-93] (Anti-Lck (phospho Y394) antibody [EPR27974-93] ab318960) at 1/1000 dilution
Lane 1: COLO 201 (human colon adenocarcinoma tumor ascitic fibroblast-like cell) treated with 200 nM Dasatinib for 24 hours whole cell lysate (untreated membrane) at 20 µg with NFDM/TBST
Lane 2: Untreated COLO 201 whole cell lysate (untreated membrane) at 20 µg with NFDM/TBST
Lane 3: Untreated COLO 201 whole cell lysate (alkaline phosphatase treated membrane) at 20 µg with NFDM/TBST
Secondary
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution
Observed band size: 60 kDa, 36 kDa
Exposure time: 59s