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.
Images
![Dot Blot - Anti-Lck (phospho Y394) antibody [EPR27974-93] - BSA and Azide free (AB318961), expandable thumbnail](https://content.abcam.com/products/images/lck-phospho-y394-antibody-epr27974-93-bsa-and-azide-free-ab318961--dot-blot-img433423.jpg/_jcr_content/renditions/webp-475.webp "Dot Blot - 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), expandable thumbnail](https://content.abcam.com/products/images/lck-phospho-y394-antibody-epr27974-93-bsa-and-azide-free-ab318961--immunohistochemistry-formalin-pfa-fixed-paraffin-embedded-sections-img433383.jpg/_jcr_content/renditions/webp-475.webp "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), expandable thumbnail](https://content.abcam.com/products/images/lck-phospho-y394-antibody-epr27974-93-bsa-and-azide-free-ab318961--immunohistochemistry-formalin-pfa-fixed-paraffin-embedded-sections-img434572.jpg/_jcr_content/renditions/webp-475.webp "Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - 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), expandable thumbnail](https://content.abcam.com/products/images/lck-phospho-y394-antibody-epr27974-93-bsa-and-azide-free-ab318961--western-blot-img433352.jpg/_jcr_content/renditions/webp-475.webp "Western blot - 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), expandable thumbnail](https://content.abcam.com/products/images/lck-phospho-y394-antibody-epr27974-93-bsa-and-azide-free-ab318961--flow-cytometry-intracellular-img433795.jpg/_jcr_content/renditions/webp-475.webp "Flow Cytometry (Intracellular) - Anti-Lck (phospho Y394) antibody [EPR27974-93] - BSA and Azide free (AB318961)")
Key facts
- Isotype
- IgG
- Host species
- Rabbit
- Storage buffer
pH: 7.2 - 7.4
Constituents: 100% PBS- Form
- Liquid
- Clonality
- Monoclonal
Immunogen
- The exact immunogen used to generate this antibody is proprietary information.
Reactivity data
Select an application| Dot | Flow Cyt (Intra) | WB | IHC-P | ICC/IF | |
|---|---|---|---|---|---|
| Human | Expected | Tested | Tested | Tested | Not recommended |
| Synthetic peptide | Tested | Not recommended | Not recommended | Not recommended | Not recommended |
Dot
TestedTested
| Species | Dilution info | Notes |
|---|---|---|
Species Synthetic peptide | Dilution info - | Notes - |
ExpectedExpected
| Species | Dilution info | Notes |
|---|---|---|
Species Human | Dilution info Use at an assay dependent concentration. | Notes - |
Flow Cyt (Intra)
TestedTested
| Species | Dilution info | Notes |
|---|---|---|
Species Human | Dilution info - | Notes - |
Not recommendedNot recommended
| Species | Dilution info | Notes |
|---|---|---|
Species Synthetic peptide | Dilution info - | Notes - |
WB
TestedTested
| Species | Dilution info | Notes |
|---|---|---|
Species Human | Dilution info - | Notes - |
Not recommendedNot recommended
| Species | Dilution info | Notes |
|---|---|---|
Species Synthetic peptide | Dilution info - | Notes - |
IHC-P
TestedTested
| Species | Dilution info | Notes |
|---|---|---|
Species Human | Dilution info - | Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
Not recommendedNot recommended
| Species | Dilution info | Notes |
|---|---|---|
Species Synthetic peptide | Dilution info - | Notes - |
ICC/IF
Not recommendedNot recommended
| Species | Dilution info | Notes |
|---|---|---|
Species Human, Synthetic peptide | 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
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 phospho Y394 antibody. Carrier free. Suitable for Dot, Flow Cyt (Intra), WB, IHC-P and reacts with Synthetic peptide, Human samples.
Key facts
- Isotype
- IgG
- Host species
- Rabbit
- Storage buffer
pH: 7.2 - 7.4
Constituents: 100% PBS- Form
- Liquid
- Clonality
- Monoclonal
- Immunogen
- The exact immunogen used to generate this antibody is proprietary information.
- Carrier free
- Yes
- Clone number
- EPR27974-93
- Purification technique
- Affinity purification Protein A
- Concentration
Storage
- Shipped at conditions
- Blue Ice
- Appropriate short-term storage conditions
- +4°C
- Appropriate long-term storage conditions
- +4°C
Notes
ab318961 is the carrier-free version of Anti-Lck (phospho Y394) antibody [EPR27974-93] ab318960.
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.
Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.
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.
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.
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
Tested
We have tested this species and application combination and it works. It is covered by our product promise.
Expected
We have not tested this specific species and application combination in-house, but expect it will work. It is covered by our product promise.
Predicted
This species and application combination has not been tested, but we predict it will work based on strong homology. However, this combination is not covered by our product promise.
Not recommended
We do not recommend this combination. It is not covered by our 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.
11 product images
![Dot Blot - Anti-Lck (phospho Y394) antibody [EPR27974-93] - BSA and Azide free (ab318961), expandable thumbnail](https://content.abcam.com/products/images/lck-phospho-y394-antibody-epr27974-93-bsa-and-azide-free-ab318961--dot-blot-img433423.jpg "Dot 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)
Lck (phospho Y394) Dot Blot staining using rabbit Anti-Lck (phospho Y394) antibody
This data was developed using Anti-Lck (phospho Y394) antibody [EPR27974-93] ab318960, the same antibody clone in a different buffer formulation.
Dot blot analysis of Lck (phospho Y394) using Anti-Lck (phospho Y394) antibody [EPR27974-93] ab318960 at 1:1000 (0.519 ug/ml) followed by a Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (Goat Anti-Rabbit IgG H&L (HRP) 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] (Anti-Lck (phospho Y394) antibody [EPR27974-93] ab318960) 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
SecondaryAll lanes: Dot Blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution
Exposure time: 180s
![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Lck (phospho Y394) antibody [EPR27974-93] - BSA and Azide free (ab318961), expandable thumbnail](https://content.abcam.com/products/images/lck-phospho-y394-antibody-epr27974-93-bsa-and-azide-free-ab318961--immunohistochemistry-formalin-pfa-fixed-paraffin-embedded-sections-img433383.jpg "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)
This data was developed using Anti-Lck (phospho Y394) antibody [EPR27974-93] ab318960, the same antibody clone in a different buffer formulation.
Immunohistochemical analysis of paraffin-embedded Human tonsil tissue labeling Lck (phospho Y394) with Anti-Lck (phospho Y394) antibody [EPR27974-93] 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 Anti-Lck (phospho Y394) antibody [EPR27974-93] ab318960 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrumentCounterstained 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
![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Lck (phospho Y394) antibody [EPR27974-93] - BSA and Azide free (ab318961), expandable thumbnail](https://content.abcam.com/products/images/lck-phospho-y394-antibody-epr27974-93-bsa-and-azide-free-ab318961--immunohistochemistry-formalin-pfa-fixed-paraffin-embedded-sections-img434572.jpg "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)
This data was developed using Anti-Lck (phospho Y394) antibody [EPR27974-93] 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 Anti-Lck (phospho Y394) antibody [EPR27974-93] 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 Anti-Lck (phospho Y394) antibody [EPR27974-93] ab318960 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrumentCounterstained 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
![Western blot - Anti-Lck (phospho Y394) antibody [EPR27974-93] - BSA and Azide free (ab318961), expandable thumbnail](https://content.abcam.com/products/images/lck-phospho-y394-antibody-epr27974-93-bsa-and-azide-free-ab318961--western-blot-img433352.jpg "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)
This data was developed using Anti-Lck (phospho Y394) antibody [EPR27974-93] ab318960, the same antibody clone in a different buffer formulation.
Low expression: TT.
In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (Anti-GAPDH antibody [EPR16891] - Loading Control ab181602) staining at 1/200000 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 at 20 µg with NFDM/TBST
Lane 2: TT (human thyroid carcinoma epithelial cell) whole cell lysate at 20 µg with NFDM/TBST
SecondaryAll 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
![Flow Cytometry (Intracellular) - Anti-Lck (phospho Y394) antibody [EPR27974-93] - BSA and Azide free (ab318961), expandable thumbnail](https://content.abcam.com/products/images/lck-phospho-y394-antibody-epr27974-93-bsa-and-azide-free-ab318961--flow-cytometry-intracellular-img433795.jpg "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)
This data was developed using Anti-Lck (phospho Y394) antibody [EPR27974-93] 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 Anti-Lck (phospho Y394) antibody [EPR27974-93] ab318960 at 1/50 dilution (0.1ug) / Green and Magenta compared with a Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) (Black and Grey) isotype control.
Goat Anti-Rabbit IgG (Alexa Fluor® 488, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081) at 1/5000 dilution was used as the secondary antibody.![Western blot - Anti-Lck (phospho Y394) antibody [EPR27974-93] - BSA and Azide free (ab318961), expandable thumbnail](https://content.abcam.com/products/images/lck-phospho-y394-antibody-epr27974-93-bsa-and-azide-free-ab318961--western-blot-img433351.jpg "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)
This data was developed using Anti-Lck (phospho Y394) antibody [EPR27974-93] 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 (Anti-GAPDH antibody [EPR16891] - Loading Control ab181602) staining at 1/200000 dilution.
In Western blot, Anti-Lck antibody - Total protein control (Anti-Lck antibody [EPR20798-107] 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
SecondaryAll 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
![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Lck (phospho Y394) antibody [EPR27974-93] - BSA and Azide free (ab318961), expandable thumbnail](https://content.abcam.com/products/images/lck-phospho-y394-antibody-epr27974-93-bsa-and-azide-free-ab318961--immunohistochemistry-formalin-pfa-fixed-paraffin-embedded-sections-img433384.jpg "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)
This data was developed using Anti-Lck (phospho Y394) antibody [EPR27974-93] ab318960, the same antibody clone in a different buffer formulation.
Immunohistochemical analysis of paraffin-embedded Human spleen tissue labeling Lck (phospho Y394) with Anti-Lck (phospho Y394) antibody [EPR27974-93] 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 Anti-Lck (phospho Y394) antibody [EPR27974-93] ab318960 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrumentCounterstained 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), expandable thumbnail](https://content.abcam.com/products/images/lck-phospho-y394-antibody-epr27974-93-bsa-and-azide-free-ab318961--flow-cytometry-intracellular-img433409.jpg "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)
This data was developed using Anti-Lck (phospho Y394) antibody [EPR27974-93] 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 Anti-Lck (phospho Y394) antibody [EPR27974-93] ab318960 at 1/50 dilution (1ug) / Magenta (Red) compared with a Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control 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, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081) at 1/5000 dilution was used as the secondary antibody.
Low expression: TT.
![Flow Cytometry (Intracellular) - Anti-Lck (phospho Y394) antibody [EPR27974-93] - BSA and Azide free (ab318961), expandable thumbnail](https://content.abcam.com/products/images/lck-phospho-y394-antibody-epr27974-93-bsa-and-azide-free-ab318961--flow-cytometry-intracellular-img433410.jpg "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)
This data was developed using Anti-Lck (phospho Y394) antibody [EPR27974-93] 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 Anti-Lck (phospho Y394) antibody [EPR27974-93] ab318960 at 1/500 dilution (0.1ug) / Magenta (Red) compared with a Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control 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, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081) at 1/5000 dilution was used as the secondary antibody.
![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Lck (phospho Y394) antibody [EPR27974-93] - BSA and Azide free (ab318961), expandable thumbnail](https://content.abcam.com/products/images/lck-phospho-y394-antibody-epr27974-93-bsa-and-azide-free-ab318961--immunohistochemistry-formalin-pfa-fixed-paraffin-embedded-sections-img433385.jpg "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)
This data was developed using Anti-Lck (phospho Y394) antibody [EPR27974-93] ab318960, the same antibody clone in a different buffer formulation.
Immunohistochemical analysis of paraffin-embedded Human breast cancer tissue labeling Lck (phospho Y394) with Anti-Lck (phospho Y394) antibody [EPR27974-93] 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 Anti-Lck (phospho Y394) antibody [EPR27974-93] ab318960 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrumentCounterstained 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
![Western blot - Anti-Lck (phospho Y394) antibody [EPR27974-93] - BSA and Azide free (ab318961), expandable thumbnail](https://content.abcam.com/products/images/lck-phospho-y394-antibody-epr27974-93-bsa-and-azide-free-ab318961--western-blot-img433353.jpg "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)
This data was developed using Anti-Lck (phospho Y394) antibody [EPR27974-93] ab318960, the same antibody clone in a different buffer formulation.
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 (Anti-Lck antibody [EPR20798-107] 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
SecondaryAll 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 (phospho Y394) antibody [EPR27974-93] - BSA and Azide free (ab318961), expandable thumbnail](https://content.abcam.com/products/images/lck-phospho-y394-antibody-epr27974-93-bsa-and-azide-free-ab318961--western-blot-img433351.jpg/_jcr_content/renditions/webp-475.webp "Western blot - 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), expandable thumbnail](https://content.abcam.com/products/images/lck-phospho-y394-antibody-epr27974-93-bsa-and-azide-free-ab318961--immunohistochemistry-formalin-pfa-fixed-paraffin-embedded-sections-img433384.jpg/_jcr_content/renditions/webp-475.webp "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), expandable thumbnail](https://content.abcam.com/products/images/lck-phospho-y394-antibody-epr27974-93-bsa-and-azide-free-ab318961--flow-cytometry-intracellular-img433409.jpg/_jcr_content/renditions/webp-475.webp "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), expandable thumbnail](https://content.abcam.com/products/images/lck-phospho-y394-antibody-epr27974-93-bsa-and-azide-free-ab318961--flow-cytometry-intracellular-img433410.jpg/_jcr_content/renditions/webp-475.webp "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), expandable thumbnail](https://content.abcam.com/products/images/lck-phospho-y394-antibody-epr27974-93-bsa-and-azide-free-ab318961--immunohistochemistry-formalin-pfa-fixed-paraffin-embedded-sections-img433385.jpg/_jcr_content/renditions/webp-475.webp "Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - 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), expandable thumbnail](https://content.abcam.com/products/images/lck-phospho-y394-antibody-epr27974-93-bsa-and-azide-free-ab318961--western-blot-img433353.jpg/_jcr_content/renditions/webp-475.webp "Western blot - Anti-Lck (phospho Y394) antibody [EPR27974-93] - BSA and Azide free (ab318961)")
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