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AB300118

Anti-Lyn (phospho Y397) antibody [EPR26354-1]

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Rabbit Recombinant Monoclonal LYN phospho Y397 antibody. Suitable for WB, Dot, ICC/IF, Flow Cyt (Intra), IP and reacts with Human samples.

View Alternative Names

JTK8, LYN, Tyrosine-protein kinase Lyn, Lck/Yes-related novel protein tyrosine kinase, V-yes-1 Yamaguchi sarcoma viral related oncogene homolog, p53Lyn, p56Lyn

6 Images
Flow Cytometry (Intracellular) - Anti-Lyn (phospho Y397) antibody [EPR26354-1] (AB300118)
  • Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Anti-Lyn (phospho Y397) antibody [EPR26354-1] (AB300118)

Flow cytometric analysis of 4% Paraformaldehyde fixed 90% Methanol permeabilized COLO 201 (Human colon adenocarcinoma) 200nM Dasatinib for 24 hours (Red) / Untreated control (Green) cells labelling Lyn (phospho Y397) with ab300118 at 1/500 compared with a Rabbit monoclonal IgG (ab172730) / Black isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody).

Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/2000 was used as the secondary antibody.

Dasatinib treatment inhibit the phosphorylation of Lyn at Y397 (PMID : 18451159).

Immunocytochemistry/ Immunofluorescence - Anti-Lyn (phospho Y397) antibody [EPR26354-1] (AB300118)
  • ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-Lyn (phospho Y397) antibody [EPR26354-1] (AB300118)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized COLO 201 (human derived from metastatic site ascites bipolar, slightly refractile, fibroblast-like cell) cells labelling Lyn (phospho Y397) with ab300118 at 1/50 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed (ab150081) antibody at 1/1000 dilution (Green).

Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) (ab195889)  was used to counterstain tubulin at 1/200 dilution. The Nuclear counterstain was DAPI (Blue).

Secondary antibody only control : Secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed (ab150081) at 1/1000 dilution.

Immunoprecipitation - Anti-Lyn (phospho Y397) antibody [EPR26354-1] (AB300118)
  • IP

Lab

Immunoprecipitation - Anti-Lyn (phospho Y397) antibody [EPR26354-1] (AB300118)

Lyn (phospho Y397) was immunoprecipitated from 0.35 mg COLO 205 whole cell lysate with ab300118 at 1/30 dilution. Western blot was performed on the immunoprecipitate using ab300118 at 1/1000 dilution.

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

All lanes:

Immunoprecipitation - Anti-Lyn (phospho Y397) antibody [EPR26354-1] (ab300118) at 1/1000 dilution

Lane 1:

COLO 205 (human colorectal adenocarcinoma cell) whole cell lysate at 10 µg

Lane 2:

ab300118 at 1/30 IP in COLO 205 whole cell lysate at 10 µg

Lane 3:

Rabbit monoclonal IgG (<a href='/en-us/products/primary-antibodies/rabbit-igg-monoclonal-epr25a-isotype-control-ab172730'>ab172730</a>) instead of ab300118 in COLO 205 whole cell lysate at 10 µg

Secondary

All lanes:

Immunoprecipitation - VeriBlot for IP Detection Reagent (HRP) (<a href='/en-us/products/reagents/veriblot-for-ip-detection-reagent-hrp-ab131366'>ab131366</a>) at 1/5000 dilution

Observed band size: 60 kDa

false

Exposure time: 15s

Western blot - Anti-Lyn (phospho Y397) antibody [EPR26354-1] (AB300118)
  • WB

Lab

Western blot - Anti-Lyn (phospho Y397) antibody [EPR26354-1] (AB300118)

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining.

Dasatinib treatment inhibits the phosphorylation of Lyn at Y397 (PMID : 18451159). The antibody may cross-react with phosphorylated HCK(Tyr411), LCK(Tyr394), BLK(Tyr389), Src(Tyr416), Fyn(Tyr420) and Yes(Tyr426).

All lanes:

Western blot - Anti-Lyn (phospho Y397) antibody [EPR26354-1] (ab300118) at 1/1000 dilution

Lane 1:

Untreated COLO 201 (human colorectal adenocarcinoma cell) whole cell lysate at 40 µg

Lane 2:

COLO 201 treated with 200 nM Dasatinib for 24 hours, whole cell lysate at 40 µg

Lane 3:

Untreated COLO 205 (human colorectal adenocarcinoma cell) whole cell lysate at 40 µg

Lane 4:

COLO 205 treated with 200 nM Dasatinib for 24 hours, whole cell lysate at 40 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: 58 kDa

false

Exposure time: 26s

Western blot - Anti-Lyn (phospho Y397) antibody [EPR26354-1] (AB300118)
  • WB

Lab

Western blot - Anti-Lyn (phospho Y397) antibody [EPR26354-1] (AB300118)

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining.

All lanes:

Western blot - Anti-Lyn (phospho Y397) antibody [EPR26354-1] (ab300118) at 1/1000 dilution

Lane 1:

COLO 205 (human colorectal adenocarcinoma cell) whole cell lysate at 20 µg

Lane 2:

COLO 205 (human colorectal adenocarcinoma cell) whole cell lysate (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: 58 kDa

false

Exposure time: 26s

Dot Blot - Anti-Lyn (phospho Y397) antibody [EPR26354-1] (AB300118)
  • Dot

Lab

Dot Blot - Anti-Lyn (phospho Y397) antibody [EPR26354-1] (AB300118)

Dot blot analysis of Lyn (phospho Y397) using ab300118 at 1/1000 followed by a Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1/100000 dilution.

This antibody cross-reacts with human YES (phospho T426), FYN (phospho T420), SRC (phospho T419), and Human LCK (phospho T394), but does not react with Human BLK (phospho T389).

All lanes:

Dot Blot - Anti-Lyn (phospho Y397) antibody [EPR26354-1] (ab300118) at 1/1000 dilution

Lane 1:

Human Lyn (phospho Y397) peptide a

Lane 2:

Human Lyn (phospho Y397) peptide b

Lane 3:

Human Lyn non-phospho peptide

Lane 4:

Human BLK (phospho T389) peptide

Lane 5:

Human YES (phospho T426)/ FYN (phospho T420)/ SRC (phospho T419) peptide

Lane 6:

Human LCK (phospho T394) peptide

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: 3min

  • Carrier free

    Anti-Lyn (phospho Y397) antibody [EPR26354-1] (BSA and Azide free)

Key facts

Host species

Rabbit

Clonality

Monoclonal

Clone number

EPR26354-1

Isotype

IgG

Carrier free

No

Reacts with

Human

Applications

WB, IP, Flow Cyt (Intra), ICC/IF, 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"}, "WB" : {"fullname" : "Western blot", "shortname":"WB"}, "IHCP" : {"fullname" : "Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)", "shortname":"IHC-P"}, "Dot" : {"fullname" : "Dot Blot", "shortname":"Dot"}, "ICCIF" : {"fullname" : "Immunocytochemistry/ Immunofluorescence", "shortname":"ICC/IF"}, "FlowCytIntra" : {"fullname" : "Flow Cytometry (Intracellular)", "shortname":"Flow Cyt (Intra)"}, "IP" : {"fullname" : "Immunoprecipitation", "shortname":"IP"} }, "product-promise": { "all": "all", "testedAndGuaranteed": "tested", "guaranteed": "expected", "predicted": "predicted", "notRecommended": "not-recommended" } }, "values": { "Human": { "WB-species-checked": "testedAndGuaranteed", "WB-species-dilution-info": "1/1000", "WB-species-notes": "<p></p>", "IHCP-species-checked": "notRecommended", "IHCP-species-dilution-info": "", "IHCP-species-notes": "<p></p>", "Dot-species-checked": "testedAndGuaranteed", "Dot-species-dilution-info": "1/1000", "Dot-species-notes": "<p></p>", "ICCIF-species-checked": "testedAndGuaranteed", "ICCIF-species-dilution-info": "1/50", "ICCIF-species-notes": "<p></p>", "FlowCytIntra-species-checked": "testedAndGuaranteed", "FlowCytIntra-species-dilution-info": "1/500", "FlowCytIntra-species-notes": "<p></p>", "IP-species-checked": "testedAndGuaranteed", "IP-species-dilution-info": "1/30", "IP-species-notes": "<p></p>" }, "Mouse": { "WB-species-checked": "notRecommended", "WB-species-dilution-info": "1/1000", "WB-species-notes": "<p></p>", "IHCP-species-checked": "notRecommended", "IHCP-species-dilution-info": "", "IHCP-species-notes": "<p></p>", "Dot-species-checked": "notRecommended", "Dot-species-dilution-info": "", "Dot-species-notes": "", "ICCIF-species-checked": "notRecommended", "ICCIF-species-dilution-info": "", "ICCIF-species-notes": "", "FlowCytIntra-species-checked": "notRecommended", "FlowCytIntra-species-dilution-info": "", "FlowCytIntra-species-notes": "", "IP-species-checked": "notRecommended", "IP-species-dilution-info": "", "IP-species-notes": "" }, "Rat": { "WB-species-checked": "notRecommended", "WB-species-dilution-info": "1/1000", "WB-species-notes": "<p></p>", "IHCP-species-checked": "notRecommended", "IHCP-species-dilution-info": "", "IHCP-species-notes": "<p></p>", "Dot-species-checked": "notRecommended", "Dot-species-dilution-info": "", "Dot-species-notes": "", "ICCIF-species-checked": "notRecommended", "ICCIF-species-dilution-info": "", "ICCIF-species-notes": "", "FlowCytIntra-species-checked": "notRecommended", "FlowCytIntra-species-dilution-info": "", "FlowCytIntra-species-notes": "", "IP-species-checked": "notRecommended", "IP-species-dilution-info": "", "IP-species-notes": "" } } }
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Product details

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.

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Properties and storage information

Form
Liquid
Purification technique
Affinity purification Protein A
Storage buffer
pH: 7.2 - 7.4 Preservative: 0.01% Sodium azide Constituents: PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
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
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Supplementary information

This supplementary information is collated from multiple sources and compiled automatically.

Lyn kinase also known as p56 or Lyn protein is a member of the Src family of protein tyrosine kinases. It possesses a molecular weight of approximately 56 kDa. Lyn is mainly expressed in hematopoietic cells such as B-lymphocytes mast cells and myeloid cells. It plays a critical role in the signaling pathways involving these cells. Lyn is located on the inner surface of the plasma membrane and requires lipid modification for membrane association. Additionally it is present in neuronal tissues and other non-immune cell types reflecting a broader role in cellular signal transduction.
Biological function summary

As a regulator in signal transduction Lyn kinase is involved in the formation of protein complexes that modulate cell activation and proliferation. It participates in the phosphorylation of downstream substrates which leads to the initiation of various cellular responses. Lyn modulates pathways involving inflammation and immune responses impacting the development and function of immune cells. It is important for balancing activation and inhibition signals in the immune system ensuring a controlled immune response.

Pathways

Lyn kinase interacts with the B-cell receptor (BCR) signaling pathway and the Fc epsilon RI signaling pathway in mast cells. It associates with other Src family kinases like Fyn and Lck to regulate BCR signaling which influences B-cell development and differentiation. Lyn also acts upstream of pathways that control the release of inflammatory mediators. Through its role in these pathways Lyn indirectly impacts downstream processes and affects cellular communication and immune system homeostasis.

Lyn dysfunction has implications for conditions like chronic myeloid leukemia and systemic lupus erythematosus. Abnormal Lyn activity alters signal transduction leading to unregulated cell proliferation and immune dysregulation. In chronic myeloid leukemia Lyn interacts with the BCR/ABL fusion protein contributing to the malignant transformation of hematopoietic cells. In systemic lupus erythematosus Lyn regulates signaling pathways that prevent autoimmunity; imbalances can lead to an excessive immune response. These connections highlight Lyn's importance in both cell regulation and its involvement in pathological states.
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Product protocols

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

Non-receptor tyrosine-protein kinase that transmits signals from cell surface receptors and plays an important role in the regulation of innate and adaptive immune responses, hematopoiesis, responses to growth factors and cytokines, integrin signaling, but also responses to DNA damage and genotoxic agents. Functions primarily as negative regulator, but can also function as activator, depending on the context. Required for the initiation of the B-cell response, but also for its down-regulation and termination. Plays an important role in the regulation of B-cell differentiation, proliferation, survival and apoptosis, and is important for immune self-tolerance. Acts downstream of several immune receptors, including the B-cell receptor, CD79A, CD79B, CD5, CD19, CD22, FCER1, FCGR2, FCGR1A, TLR2 and TLR4. Plays a role in the inflammatory response to bacterial lipopolysaccharide. Mediates the responses to cytokines and growth factors in hematopoietic progenitors, platelets, erythrocytes, and in mature myeloid cells, such as dendritic cells, neutrophils and eosinophils. Acts downstream of EPOR, KIT, MPL, the chemokine receptor CXCR4, as well as the receptors for IL3, IL5 and CSF2. Plays an important role in integrin signaling. Regulates cell proliferation, survival, differentiation, migration, adhesion, degranulation, and cytokine release. Involved in the regulation of endothelial activation, neutrophil adhesion and transendothelial migration (PubMed : 36932076). Down-regulates signaling pathways by phosphorylation of immunoreceptor tyrosine-based inhibitory motifs (ITIM), that then serve as binding sites for phosphatases, such as PTPN6/SHP-1, PTPN11/SHP-2 and INPP5D/SHIP-1, that modulate signaling by dephosphorylation of kinases and their substrates. Phosphorylates LIME1 in response to CD22 activation. Phosphorylates BTK, CBL, CD5, CD19, CD72, CD79A, CD79B, CSF2RB, DOK1, HCLS1, LILRB3/PIR-B, MS4A2/FCER1B, SYK and TEC. Promotes phosphorylation of SIRPA, PTPN6/SHP-1, PTPN11/SHP-2 and INPP5D/SHIP-1. Mediates phosphorylation of the BCR-ABL fusion protein. Required for rapid phosphorylation of FER in response to FCER1 activation. Mediates KIT phosphorylation. Acts as an effector of EPOR (erythropoietin receptor) in controlling KIT expression and may play a role in erythroid differentiation during the switch between proliferation and maturation. Depending on the context, activates or inhibits several signaling cascades. Regulates phosphatidylinositol 3-kinase activity and AKT1 activation. Regulates activation of the MAP kinase signaling cascade, including activation of MAP2K1/MEK1, MAPK1/ERK2, MAPK3/ERK1, MAPK8/JNK1 and MAPK9/JNK2. Mediates activation of STAT5A and/or STAT5B. Phosphorylates LPXN on 'Tyr-72'. Kinase activity facilitates TLR4-TLR6 heterodimerization and signal initiation. Phosphorylates SCIMP on 'Tyr-107'; this enhances binding of SCIMP to TLR4, promoting the phosphorylation of TLR4, and a selective cytokine response to lipopolysaccharide in macrophages (By similarity). Phosphorylates CLNK (By similarity). Phosphorylates BCAR1/CAS and NEDD9/HEF1 (PubMed : 9020138).
See full target information LYN phospho Y397
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Product promise

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