Phospho-SRC (Y216) Antibody
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Product Specs
Buffer
Liquid in PBS containing 50% glycerol, 0.5% BSA and 0.02% sodium azide.
Form
Liquid
Lead Time
Typically, we can ship your orders within 1-3 business days of receipt. Delivery times may vary depending on the purchase method or location. For specific delivery times, please contact your local distributors.
Synonyms
ASV antibody; Avian sarcoma virus antibody; AW259666 antibody; c SRC antibody; CDNA FLJ14219 fis clone NT2RP3003800 highly similar to Rattus norvegicus tyrosine protein kinase pp60 c src mRNA antibody; cSrc antibody; EC 2.7.10.2 antibody; Neuronal CSRC tyrosine specific protein kinase antibody; Neuronal proto-oncogene tyrosine-protein kinase Src antibody; Neuronal SRC antibody; Oncogene SRC antibody; OTTHUMP00000174476 antibody; OTTHUMP00000174477 antibody; p60 Src antibody; p60-Src antibody; p60c-src antibody; p60Src antibody; pp60c src antibody; pp60c-src antibody; pp60csrc antibody; Proto oncogene tyrosine protein kinase Src antibody; Proto-oncogene c-Src antibody; Proto-oncogene tyrosine-protein kinase Src antibody; Protooncogene SRC antibody; Protooncogene SRC Rous sarcoma antibody; Src antibody; SRC Oncogene antibody; SRC proto oncogene non receptor tyrosine kinase antibody; SRC_HUMAN antibody; SRC1 antibody; Tyrosine kinase pp60c src antibody; Tyrosine protein kinase SRC 1 antibody; Tyrosine protein kinase SRC1 antibody; v src avian sarcoma (Schmidt Ruppin A2) viral oncogene homolog antibody; V src sarcoma (Schmidt Ruppin A 2) viral oncogene homolog (avian) antibody; v src sarcoma (Schmidt Ruppin A 2) viral oncogene homolog avian antibody
Target Names
Uniprot No.
Target Background
Function
Src is a non-receptor protein tyrosine kinase that is activated upon engagement of various cellular receptors. These receptors include immune response receptors, integrins and other adhesion receptors, receptor protein tyrosine kinases, G protein-coupled receptors, and cytokine receptors. Src participates in signaling pathways that regulate a wide range of biological processes, including gene transcription, immune response, cell adhesion, cell cycle progression, apoptosis, migration, and transformation. Due to functional redundancy among members of the Src kinase family, pinpointing the specific role of each Src kinase is challenging. Src appears to be a primary kinase activated following receptor engagement and plays a role in the activation of other protein tyrosine kinase (PTK) families. Receptor clustering or dimerization leads to Src recruitment to the receptor complexes, where it phosphorylates tyrosine residues within the receptor cytoplasmic domains. Src plays a crucial role in regulating cytoskeletal organization through phosphorylation of specific substrates such as AFAP1. Phosphorylation of AFAP1 allows the Src SH2 domain to bind to AFAP1 and localize to actin filaments. Cytoskeletal reorganization is also regulated through the phosphorylation of cortactin (CTTN) (Probable). When cells adhere via focal adhesions to the extracellular matrix, signals are transmitted by integrins into the cell, resulting in tyrosine phosphorylation of various focal adhesion proteins, including PTK2/FAK1 and paxillin (PXN). In addition to phosphorylating focal adhesion proteins, Src is also active at cell-cell contact sites (adherens junctions) and phosphorylates substrates such as beta-catenin (CTNNB1), delta-catenin (CTNND1), and plakoglobin (JUP). Another type of cell-cell junction, the gap junction, is also a target for Src, which phosphorylates connexin-43 (GJA1). Src is implicated in the regulation of pre-mRNA processing and phosphorylates RNA-binding proteins such as KHDRBS1 (Probable). Src also plays a role in PDGF-mediated tyrosine phosphorylation of both STAT1 and STAT3, leading to increased DNA binding activity of these transcription factors. Src is involved in the RAS pathway through phosphorylation of RASA1 and RASGRF1. It plays a role in EGF-mediated calcium-activated chloride channel activation. Src is required for epidermal growth factor receptor (EGFR) internalization through phosphorylation of clathrin heavy chain (CLTC and CLTCL1) at 'Tyr-1477'. Src is involved in beta-arrestin (ARRB1 and ARRB2) desensitization through phosphorylation and activation of GRK2, leading to beta-arrestin phosphorylation and internalization. Src has a critical role in stimulating the CDK20/MAPK3 mitogen-activated protein kinase cascade by epidermal growth factor (Probable). Src may not only mediate the transduction of mitogenic signals at the plasma membrane but also control progression through the cell cycle via interaction with regulatory proteins in the nucleus. Src plays a crucial role in osteoclastic bone resorption in conjunction with PTK2B/PYK2. Both the formation of a Src-PTK2B/PYK2 complex and Src kinase activity are necessary for this function. Src is recruited to activated integrins by PTK2B/PYK2, thereby phosphorylating CBL, which in turn induces the activation and recruitment of phosphatidylinositol 3-kinase to the cell membrane in a signaling pathway that is critical for osteoclast function. Src promotes energy production in osteoclasts by activating mitochondrial cytochrome C oxidase. It phosphorylates DDR2 on tyrosine residues, thereby promoting its subsequent autophosphorylation. Src phosphorylates RUNX3 and COX2 on tyrosine residues, TNK2 on 'Tyr-284', and CBL on 'Tyr-731'. Src enhances DDX58/RIG-I-elicited antiviral signaling. Src phosphorylates PDPK1 at 'Tyr-9', 'Tyr-373', and 'Tyr-376'. It phosphorylates BCAR1 at 'Tyr-128'. Src phosphorylates CBLC at multiple tyrosine residues, phosphorylation at 'Tyr-341' activates CBLC E3 activity. Src is involved in anchorage-independent cell growth. Src is required for podosome formation. Src mediates IL6 signaling by activating the YAP1-NOTCH pathway to induce inflammation-induced epithelial regeneration.
Gene References Into Functions
- Mutation in c-Src phosphorylation site of either HK1 or HK2 remarkably abrogates the stimulating effects of c-Src on glycolysis, cell proliferation, migration, invasion, tumorigenesis and metastasis PMID: 28054552
- Results showed that CAV-1 could promote anchorage-independent growth and anoikis resistance in detached SGC-7901 cells, which was associated with the activation of Src-dependent epidermal growth factor receptor-integrin beta signaling as well as the phosphorylation of PI3K/Akt and MEK/ERK signaling pathways PMID: 30088837
- This study shows that Leu33Pro polymorphism of integrin beta 3 modulates platelet Src pY418 and focal adhesion kinase pY397 phosphorylation in response to abnormally high shear stress. While physiological shear stress does not affect platelet signaling, abnormally high shear stress considerably elevates Src and FAK phosphorylation in both Pro33 and Leu33 platelets. PMID: 29965811
- High SRC expression is associated with lung adenocarcinoma. PMID: 30015929
- While activation in c-Src is strictly controlled by ATP-binding and phosphorylation, the authors find that activating conformational transitions are spontaneously sampled in Hsp90-dependent Src mutants. PMID: 28290541
- High SRC expression is associated with gastric cancer cell migration. PMID: 30015970
- Src kinase mediates UV-induced TRPV1 trafficking into the cell membrane in HaCaT keratinocytes. PMID: 29080357
- Src kinase activation by nitric oxide promotes resistance to anoikis in tumor cell lines. PMID: 29651879
- Src and Aurora-A interact upon Golgi ribbon fragmentation; Src phosphorylates Aurora-A at tyrosine 148, and this specific phosphorylation is required for Aurora-A localization at the centrosomes. PMID: 27242098
- Study demonstrated that c-Src contributed to hypoxic microenvironment-rendered paclitaxel resistance in human epithelial ovarian cancer cells by G2/M phase arrest deterioration, and through c-Src suppression, FV-429 was capable of reversing the resistance by blocking the c-Src/Stat3/HIF-1alpha pathway. PMID: 29324735
- Data demonstrated that the Src/Fn14/NF-kappaB axis plays a critical role in NSCLC metastasis. PMID: 29500337
- Results suggest that Src promotes EGF-stimulated EMT and migration by upregulation of ZEB1 and ZEB2 through the AKT signaling pathway in gastric cancer cells. PMID: 29052277
- Combined targeting of AKT and SRC resulted in synergistic efficacy against human pancreatic cancer growth and metastasis. PMID: 29978609
- Important roles for c-Src tyrosine kinase in phosphorylation and activation of SLC11A1 in macrophages PMID: 29723216
- Our data suggest that targeting Src signaling may be an effective approach to the treatment of ALK-non-small cell lung cancer (NSCLC) with acquired resistance to ALK inhibitors. PMID: 29048652
- Src kinase in chemo-naive human primary osteosarcoma cells is differentially activated. PMID: 28786551
- This study demonstrates that simultaneous inhibition of c-Met and Src signaling in MD-MSCs triggers apoptosis and reveals vulnerable pathways that could be exploited to develop NF2 therapies. PMID: 28775147
- Syntenin mediates SRC function in exosomal cell-to-cell communication. PMID: 29109268
- Endothelial cell-derived matrix promotes the metabolic functional maturation of hepatocyte via integrin-Src signaling. PMID: 28470937
- The expression of Src under the influence of nilotinib, dasatinib, erlotinib, gefitinib, and afatinib was studied in HPV-positive head and neck squamous cell carcinomas. Src expression was significantly increased by all tested tyrosine kinase inhibitors. PMID: 29715092
- Multivariate Cox regression analysis suggested that PTPRA expression was an independent prognostic factor in SCC patients. In the cellular models, PTPRA promotes SCC cell proliferation through modulating Src activation as well as cell cycle progression. In conclusion, higher PTPRA level was associated with worse prognosis of SCC patients and PTPRA could promote the cell cycle progression PMID: 28656243
- The c-Src/MAPK/NF-kB signaling pathway may contribute to the pathogenesis of pre-eclampsia PMID: 28544129
- Data indicate the role of tyrosine kinase c-Src (Src) in rescuing Taz (transcriptional coactivator with PDZ-binding motif) from E3 ligase SCF(beta-TrCP)-mediated degradation. PMID: 28154141
- Data suggest that response of bronchial epithelial cells to environmental carcinogen benzo[a]pyrene includes activation of AhR/Src/ERK signaling, CYP1A1 induction, and formation of stable DNA adducts. (AhR = aryl hydrocarbon receptor; Src = Src proto-oncogene kinase; ERK = extracellular signal-regulated kinases; CYP1A1 = cytochrome P450 family 1 subfamily A member 1) PMID: 29545172
- It is unclear if we may have seen greater clinical activity if we were able to fully inhibit Src in this study, but given the requirement that enrolling patients have documented disease progression on cetuximab, acquired resistant KRAS-mutant clones may have been present, limiting future strategies to reverse EGFR resistance PMID: 28280091
- This study shows that simultaneous deactivation of FAK and Src improves the pathology of hypertrophic scar PMID: 27181267
- Mutations in the germline and somatic DNA of the TEK gene were identified and analyze the expression level of Src and phospho-Src (p-Src) in tumor and healthy tissues from patients with facial cutaneo-mucosal venous malformations. PMID: 28316284
- SOCS1 antagonizes epithelial-mesenchymal transition by suppressing Src activity, leading to thioredoxin expression and down-regulation of ROS levels in colon cancer cells PMID: 27613835
- These findings suggest that the integrin beta4-FAK/Src signaling axis may play a crucial role in clonorchiasis-associated cholangiocarcinoma metastasis during tumor progression. PMID: 28286026
- Estrogen receptor-Src signaling plays an important role in ER (+) breast cancer, which shows a high potential for bone metastasis. PMID: 28472954
- Thrombin binding to PAR-1 receptor activated Gi-protein/c-Src/Pyk2/EGFR/PI3K/Akt/p42/p44 MAPK cascade, which in turn elicited AP-1 activation and ultimately evoked MMP-9 expression and cell migration in SK-N-SH cells. PMID: 27181591
- Whereas Src activation under shear stress is dominantly ligand-dependent, FAK signaling seems to be mostly shear induced. PMID: 27467982
- We provide evidence here that Rab7 is a substrate of Src kinase, and is tyrosine-phosphorylated by Src, with Y183 residue of Rab7 being the optimal phosphorylation site for Src. Further investigations demonstrated that the tyrosine phosphorylation of Rab7 depends on the guanine nucleotide binding activity of Rab7 and the activity of Src kinase. PMID: 28336235
- Expression of LINC00520 is regulated by oncogenic Src, PIK3CA and STAT3, and may contribute to the molecular etiology of breast cancer. PMID: 27626181
- Findings indicate the importance of Src-Stat3 signaling cascade in gallic acid (GA)-mediated tumor-suppression activity and a therapeutic insight of GA for acquired resistance to EGF receptor tyrosine kinase inhibitors in lung cancer. PMID: 27419630
- Memo facilitates ER-alpha and c-Src interaction, ER-alpha Y537 phosphorylation, and has the ability to control ER-alpha extra-nuclear localization in breast cancer cells. PMID: 27472465
- Data show that MLLT11/AF1q-induced PDGFR signaling enhanced STAT3 activity through Src kinase activation. PMID: 27259262
- Loss of myristoylation abolished the tumorigenic potential of Src and its synergy with androgen receptor in mediating tumor invasion. PMID: 29038344
- N-WASP positively regulates demarcation membrane system development and proplatelet formation, and the Src family kinases in association with CDC42 regulate proplatelet formation through N-WASP PMID: 27685868
- Phosphorylation of mATG9 at Tyr8 by Src and at Ser14 by ULK1 functionally cooperate to promote interactions between mATG9 and the AP1/2 complex. PMID: 27934868
- Data suggest that myristoylation of Src kinase is essential to facilitate Src-induced and high-fat diet-accelerated prostatic tumor progression; targeting Src kinase myristoylation, which is required for Src kinase association at cellular membrane, blocks dietary fat-accelerated tumorigenesis. PMID: 28939770
- Elevated levels of cellular Src in serum and phosphorylated Src in primary nasopharyngeal carcinoma tissue correlated with poor outcomes of these patients PMID: 27078847
- Results indicate that src-family kinase (Src) is an upstream kinase of T-LAK cell-originated protein kinase (TOPK). PMID: 27016416
- We suggest that the induction of SRC results in increased prostate cancer metastasis that is linked to the dysregulation of the AR signaling pathway through the inactivation of miR-203 PMID: 27028864
- Data show that afatinib resistant clones were selectively killed by knock down of ERBB3 + c-MET + c-KIT, but not by the individual or doublet knock down combinations, and the combination of afatinib with the SRC family inhibitor dasatinib killed afatinib resistant H1975 cells in a greater than additive fashion. PMID: 26934000
- These results suggest that stabilization of delta-catenin by Hakai is dependent on Src. PMID: 28069439
- The protein kinase activity of PI3K phosphorylates serine residue 70 on Src to enhance its activity and induce EGFR transactivation following betaAR stimulation. PMID: 27169346
- Data show that the solubilising factor UNC119 sequesters myristoylated Src family protein tyrosine kinases (SFKs) to maintain its enrichment at the plasma membrane to enable signal transduction. PMID: 28740133
- Data indicate a role for AXL receptor tyrosine kinase (AXL) in regulating the nuclear translocation of epidermal growth factor receptor (EGFR) and suggest that AXL-mediated SRC family kinases (SFKs) and neuregulin-1 (NRG1) expression promote this process. PMID: 28049763
- High Src expression is associated with breast cancer. PMID: 28754671
Database Links
Involvement In Disease
Thrombocytopenia 6 (THC6)
Protein Families
Protein kinase superfamily, Tyr protein kinase family, SRC subfamily
Subcellular Location
Cell membrane; Lipid-anchor. Mitochondrion inner membrane. Nucleus. Cytoplasm, cytoskeleton. Cytoplasm, perinuclear region. Cell junction, focal adhesion.
Tissue Specificity
Expressed ubiquitously. Platelets, neurons and osteoclasts express 5-fold to 200-fold higher levels than most other tissues.
Q&A
What experimental challenges arise when using Phospho-SRC (Y216) Antibody in complex signaling pathway studies?
Phospho-SRC (Y216) antibodies are critical for studying Src kinase activation and signaling cascades, but their application poses challenges:
Cross-Reactivity Concerns
Src family kinases (SFKs) share conserved phosphorylation sites, risking off-target detection. For example, Yes and Fyn kinases may phosphorylate homologous Y216/Y222 residues, requiring validation with specific inhibitors or knockout models .
Dynamic Phosphorylation Kinetics
Y216 phosphorylation is transient and context-dependent. In prostate cancer models, EGF stimulation rapidly induces Y216-GSK-3 phosphorylation, necessitating precise timing in sample collection .
Optimization in Multi-Assay Platforms
How can researchers validate the specificity of Phospho-SRC (Y216) Antibody in their experimental system?
Validation requires multi-tiered approaches:
**1. Peptide Competition Assays
Incubate antibody with phospho-Y216 peptide vs. non-phosphorylated control. A >50% reduction in signal confirms epitope specificity .
**2. Knockout/Overexpression Models
Use SYF cells (lacking c-Src, Yes, Fyn) or reconstitute with Y216F mutants to eliminate background signal. Reconstitution with CA-Src should restore phosphorylation .
**3. Cross-Species Reactivity Testing
Confirm reactivity with human, mouse, and rat homologs through sequence alignment and immunoblotting .
What data contradictions might occur when correlating Phospho-SRC (Y216) levels with functional outcomes?
Discrepancies often stem from:
**A. Compartmentalization Effects
Y216 phosphorylation may localize to specific subcellular regions (e.g., cell membrane vs. cytoplasm), requiring imaging-based validation .
**B. Redundant Kinase Activity
In prostate cancer, Src-mediated Y216-GSK-3 phosphorylation may coexist with Akt-dependent Ser9/21-GSK-3 inhibition, complicating interpretation .
**C. Drug-Induced Feedback Loops
Dasatinib treatment initially increases Akt phosphorylation (S473) while suppressing Y216-GSK-3, necessitating time-course analyses .
How does Phospho-SRC (Y216) relate to drug resistance mechanisms in cancer?
Y216 phosphorylation enhances Src kinase activity, contributing to adaptive resistance:
Key Findings
Therapeutic Implications
Dasatinib inhibits Y216-GSK-3 phosphorylation, reducing tumor xenograft growth, but requires combinatorial strategies to mitigate feedback signaling .
What advanced techniques are recommended for quantifying Phospho-SRC (Y216) in clinical samples?
Quantification demands precision due to low-abundance phosphorylation:
**1. Mass Spectrometry-Based Phosphoproteomics
Shotgun MS with phosphotyrosine enrichment can identify Y216-Src and homologous sites (e.g., Y222-Yes) in patient-derived organoids .
**2. Flow Cytometry with Isotype Controls
Use mononuclear cell gating and geometric mean fluorescence ratios to normalize variability. Example workflow:
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Fixation: 4% PFA for 10 min.
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Permeabilization: Methanol at -20°C overnight.
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Staining: Primary antibody (1:1000), secondary FITC-conjugated .
How do researchers address antibody lot-to-lot variability in longitudinal studies?
Standardization protocols minimize variability:
**A. Titration Experiments
Perform dose-response curves for each new lot using pervanadate-treated HepG2 cells (positive control) and untreated samples (negative control) .
**B. Positive Controls
Include phosphorylated Src peptides or recombinant Y216-phosphorylated Src in each experiment .
What alternative approaches exist for studying Src Y216 phosphorylation when antibody availability is limited?
Alternative methods include:
**1. Kinase Activity Assays
Use Src-specific substrates (e.g., c-Abl peptide) and measure ATP hydrolysis or phosphotransfer efficiency .
**2. CRISPR-Cas9 Editing
Generate Y216F knock-in cell lines to directly assess phosphorylation dependency in signaling pathways .
**3. Proximity Ligation Assays (PLA)
Detect Y216 phosphorylation in situ with dual probes, enabling spatial resolution in tumor tissues .
How does Y216 phosphorylation differ mechanistically from Y419/Y530 phosphorylation in Src activation?
Sequential phosphorylation events drive Src activation:
What computational tools aid in interpreting Phospho-SRC (Y216) data in systems biology contexts?
Integrate phosphorylation data with pathway analysis:
**1. Network Mapping
Use tools like STRING or Cytoscape to model Y216-Src interactions with GSK-3, FAK, or RET kinases .
**2. Kinase Inhibitor Profiling
Apply dasatinib dose-response curves to predict Y216-GSK-3 inhibition thresholds in patient-derived organoids .
How does Y216 phosphorylation influence Src’s substrate specificity?
Y216 phosphorylation alters Src’s substrate preference:
Key Observations
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