AB318961

Anti-Lck (phospho Y394) antibody [EPR27974-93] - BSA and Azide free

BOND RX™ Validated
Recombinant
RabMAb
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Rabbit Recombinant Monoclonal LCK phospho Y394 antibody. Carrier free. Suitable for Dot, Flow Cyt (Intra), WB, IHC-P and reacts with Synthetic peptide, Human samples.

View 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

11 Images

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Lck (phospho Y394) antibody [EPR27974-93] - BSA and Azide free (AB318961)

IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Lck (phospho Y394) antibody [EPR27974-93] - BSA and Azide free (AB318961)

This data was developed using ab318960, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Untreated COLO 201 (human colon adenocarcinoma tumor ascitic fibroblast-like cell) cell pellets and COLO 201 treated with 200 nM Dasatinib for 24 hours labeling Lck (phospho Y394) with ab318960 at 1 : 5000 (0.104 μg/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining in (A) Untreated COLO 201 (human colon adenocarcinoma tumor ascitic fibroblast-like cell) cell pellets, and no signal was detected in (B) COLO 201 treated with 200 nM Dasatinib for 24 hours.
The section was incubated with ab318960 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument

Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

Flow Cytometry (Intracellular) - Anti-Lck (phospho Y394) antibody [EPR27974-93] - BSA and Azide free (AB318961)

Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Anti-Lck (phospho Y394) antibody [EPR27974-93] - BSA and Azide free (AB318961)

This data was developed using ab318960, the same antibody clone in a different buffer formulation.

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized COLO 205 (human colon adenocarcinoma tumor ascitic epithelial cell) (Right) / TT (human thyroid carcinoma epithelial cell) (Left) cells labelling Lck (phospho Y394) with ab318960 at 1/50 dilution (1ug) / Magenta (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue).

Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/5000 dilution was used as the secondary antibody.

Low expression : TT.

IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Lck (phospho Y394) antibody [EPR27974-93] - BSA and Azide free (AB318961)

This data was developed using ab318960, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Human spleen tissue labeling Lck (phospho Y394) with ab318960 at 1/100 (5.19 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining in human spleen without alkaline phosphatase treatment; No signal was detected when tissues were treated with alkaline phosphatase.
The section was incubated with ab318960 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument

Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Lck (phospho Y394) antibody [EPR27974-93] - BSA and Azide free (AB318961)

This data was developed using ab318960, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Human tonsil tissue labeling Lck (phospho Y394) with ab318960 at 1/100 (5.19 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining in human tonsil without alkaline phosphatase treatment; No signal was detected when tissues were treated with alkaline phosphatase.
The section was incubated with ab318960 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument

Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Lck (phospho Y394) antibody [EPR27974-93] - BSA and Azide free (AB318961)

This data was developed using ab318960, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Human breast cancer tissue labeling Lck (phospho Y394) with ab318960 at 1/100 (5.19 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining in human breast cancer without alkaline phosphatase treatment; No signal was detected when tissues were treated with alkaline phosphatase.
The section was incubated with ab318960 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument

Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Anti-Lck (phospho Y394) antibody [EPR27974-93] - BSA and Azide free (AB318961)

This data was developed using ab318960, the same antibody clone in a different buffer formulation.

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized COLO 205 (human colon adenocarcinoma tumor ascitic epithelial cell) treated with alkaline phosphatase (Left) / Untreated COLO 205 (Right) cells labelling Lck (phospho Y394) with ab318960 at 1/500 dilution (0.1ug) / Magenta (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue).

Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/5000 dilution was used as the secondary antibody.

Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Anti-Lck (phospho Y394) antibody [EPR27974-93] - BSA and Azide free (AB318961)

This data was developed using ab318960, the same antibody clone in a different buffer formulation. Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized COLO 201 (human colon adenocarcinoma tumor ascitic fibroblast-like cell) treated with 200nM Dasatinib for 24 hours (Green and Black) / Untreated COLO 201 (Magenta and Grey) cells labelling Lck (phospho Y394) with ab318960 at 1/50 dilution (0.1ug) / Green and Magenta compared with a Rabbit monoclonal IgG (ab172730) (Black and Grey) isotype control. Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/5000 dilution was used as the secondary antibody.

Supplier Data

Western blot - Anti-Lck (phospho Y394) antibody [EPR27974-93] - BSA and Azide free (AB318961)

This data was developed using ab318960, the same antibody clone in a different buffer formulation.

Low expression : TT.

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

All lanes:

Western blot - Anti-Lck (phospho Y394) antibody [EPR27974-93] (<a href='/en-us/products/primary-antibodies/lck-phospho-y394-antibody-epr27974-93-ab318960'>ab318960</a>) at 1/1000 dilution

Lane 1:

COLO 205 (human colon adenocarcinoma tumor ascitic epithelial cell) whole cell lysate at 20 µg

Lane 2:

TT (human thyroid carcinoma epithelial cell) whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Observed band size: 60 kDa,36 kDa

false

Exposure time: 180s

Supplier Data

Western blot - Anti-Lck (phospho Y394) antibody [EPR27974-93] - BSA and Azide free (AB318961)

This data was developed using ab318960, the same antibody clone in a different buffer formulation.

The expression of Lck (phospho Y394) is downregulated in response to Dasatinib treatment.

In Western blot, 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] (<a href='/en-us/products/primary-antibodies/lck-phospho-y394-antibody-epr27974-93-ab318960'>ab318960</a>) 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

Lane 2:

Untreated COLO 201 whole cell lysate (untreated membrane) at 20 µg

Lane 3:

Untreated COLO 201 whole cell lysate (alkaline phosphatase treated membrane) at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Observed band size: 60 kDa,36 kDa

false

Exposure time: 59s

Supplier Data

Western blot - Anti-Lck (phospho Y394) antibody [EPR27974-93] - BSA and Azide free (AB318961)

This data was developed using ab318960, the same antibody clone in a different buffer formulation.

In Western blot, 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] (<a href='/en-us/products/primary-antibodies/lck-phospho-y394-antibody-epr27974-93-ab318960'>ab318960</a>) at 1/1000 dilution

Lane 1:

COLO 205 (human colon adenocarcinoma tumor ascitic epithelial cell) whole cell lysate (untreated membrane) at 20 µg

Lane 2:

COLO 205 whole cell lysate (alkaline phosphatase treated membrane) at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Observed band size: 60 kDa,36 kDa

false

Exposure time: 180s

Supplier Data

Dot Blot - Anti-Lck (phospho Y394) antibody [EPR27974-93] - BSA and Azide free (AB318961)

This data was developed using ab318960, the same antibody clone in a different buffer formulation.

Dot blot analysis of Lck (phospho Y394) using ab318960 at 1 : 1000 (0.519 ug/ml) followed by a Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1 : 100,000 dilution.

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

All lanes:

Dot Blot - Anti-Lck (phospho Y394) antibody [EPR27974-93] (<a href='/en-us/products/primary-antibodies/lck-phospho-y394-antibody-epr27974-93-ab318960'>ab318960</a>) at 1/1000 dilution

Lane 1:

Lck (phospho Y394) peptide a

Lane 2:

Lck (phospho Y394) peptide b

Lane 3:

Lck non-phospho peptide c

Secondary

All lanes:

Dot Blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

false

Exposure time: 180s

Unconjugated

Anti-Lck (phospho Y394) antibody [EPR27974-93]

Key facts

Host species

Rabbit

Clonality

Monoclonal

Clone number

EPR27974-93

Isotype

IgG

Carrier free

Yes

Reacts with

Human

Applications

WB, IHC-P, Flow Cyt (Intra), Dot

applications

Immunogen

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

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Reactivity data

{ "title": "Reactivity Data", "filters": { "stats": ["", "Species", "Dilution Info", "Notes"], "tabs": { "all-applications": {"fullname" : "All Applications", "shortname": "All Applications"}, "Dot" : {"fullname" : "Dot Blot", "shortname":"Dot"}, "FlowCytIntra" : {"fullname" : "Flow Cytometry (Intracellular)", "shortname":"Flow Cyt (Intra)"}, "WB" : {"fullname" : "Western blot", "shortname":"WB"}, "IHCP" : {"fullname" : "Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)", "shortname":"IHC-P"}, "ICCIF" : {"fullname" : "Immunocytochemistry/ Immunofluorescence", "shortname":"ICC/IF"} }, "product-promise": { "all": "all", "testedAndGuaranteed": "tested", "guaranteed": "expected", "predicted": "predicted", "notRecommended": "not-recommended" } }, "values": { "Human": { "Dot-species-checked": "guaranteed", "Dot-species-dilution-info": "", "Dot-species-notes": "", "FlowCytIntra-species-checked": "testedAndGuaranteed", "FlowCytIntra-species-dilution-info": "", "FlowCytIntra-species-notes": "", "WB-species-checked": "testedAndGuaranteed", "WB-species-dilution-info": "", "WB-species-notes": "", "IHCP-species-checked": "testedAndGuaranteed", "IHCP-species-dilution-info": "", "IHCP-species-notes": "", "ICCIF-species-checked": "notRecommended", "ICCIF-species-dilution-info": "", "ICCIF-species-notes": "" }, "Synthetic peptide": { "Dot-species-checked": "testedAndGuaranteed", "Dot-species-dilution-info": "", "Dot-species-notes": "", "FlowCytIntra-species-checked": "notRecommended", "FlowCytIntra-species-dilution-info": "", "FlowCytIntra-species-notes": "", "WB-species-checked": "notRecommended", "WB-species-dilution-info": "", "WB-species-notes": "", "IHCP-species-checked": "notRecommended", "IHCP-species-dilution-info": "", "IHCP-species-notes": "", "ICCIF-species-checked": "notRecommended", "ICCIF-species-dilution-info": "", "ICCIF-species-notes": "" } } }

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Product details

ab318961 is the carrier-free version of ab318960.

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

What are the advantages of a recombinant monoclonal antibody?
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.

Conjugation ready
Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.

Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with 1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.

Compatibility
This product is compatible with the Maxpar^® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar^® is a trademark of Fluidigm Canada Inc.

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Properties and storage information

Form

Liquid

Purification technique

Affinity purification Protein A

Storage buffer

pH: 7.2 - 7.4 Constituents: PBS

Shipped at conditions

Blue Ice

Appropriate short-term storage conditions

+4°C

Appropriate long-term storage conditions

+4°C

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Supplementary information

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.

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Product protocols

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

Visit the General protocols
Visit the Troubleshooting

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Target data

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 (PubMed : 2470098). Plays a key role in T-cell antigen receptor (TCR)-linked signal transduction pathways (PubMed : 2470098). Constitutively associated with the cytoplasmic portions of the CD4 and CD8 surface receptors (PubMed : 2470098). 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 (PubMed : 2470098). 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 (PubMed : 2470098, PubMed : 40592325). 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).

See full target information Tyrosine-protein kinase Lck phospho Y394

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Product promise

We are committed to supporting your work with high-quality reagents, and we're here for you every step of the way. In the unlikely event that one of our products does not perform as expected, you're protected by our Product Promise.

For full details, please see our Terms & Conditions

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

Anti-Lck (phospho Y394) antibody [EPR27974-93] - BSA and Azide free

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Lck (phospho Y394) antibody [EPR27974-93] - BSA and Azide free (AB318961)

Flow Cytometry (Intracellular) - Anti-Lck (phospho Y394) antibody [EPR27974-93] - BSA and Azide free (AB318961)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Lck (phospho Y394) antibody [EPR27974-93] - BSA and Azide free (AB318961)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Lck (phospho Y394) antibody [EPR27974-93] - BSA and Azide free (AB318961)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Lck (phospho Y394) antibody [EPR27974-93] - BSA and Azide free (AB318961)

Flow Cytometry (Intracellular) - Anti-Lck (phospho Y394) antibody [EPR27974-93] - BSA and Azide free (AB318961)

Flow Cytometry (Intracellular) - Anti-Lck (phospho Y394) antibody [EPR27974-93] - BSA and Azide free (AB318961)

Western blot - Anti-Lck (phospho Y394) antibody [EPR27974-93] - BSA and Azide free (AB318961)

Western blot - Anti-Lck (phospho Y394) antibody [EPR27974-93] - BSA and Azide free (AB318961)

Western blot - Anti-Lck (phospho Y394) antibody [EPR27974-93] - BSA and Azide free (AB318961)

Dot Blot - Anti-Lck (phospho Y394) antibody [EPR27974-93] - BSA and Azide free (AB318961)

Related conjugates and formulations (1)

Key facts

Host species

Clonality

Clone number

Isotype

Carrier free

Reacts with

Applications

Immunogen

Complementary Products

Assay Kits

Secondary Antibodies

Primary Antibodies

Primary Antibodies

Primary Antibodies

Reactivity data

Product details

Properties and storage information

Form

Purification technique

Storage buffer

Shipped at conditions

Appropriate short-term storage conditions

Appropriate long-term storage conditions

Supplementary information

Biological function summary

Pathways

Product protocols

Target data

Product promise