Product: Phospho-Src (Tyr419) Antibody
Catalog: AF3162
Description: Rabbit polyclonal antibody to Phospho-Src (Tyr419)
Application: WB IHC IF/ICC
Cited expt.: WB
Reactivity: Human, Mouse, Rat
Prediction: Pig, Zebrafish, Bovine, Horse, Sheep, Rabbit, Dog, Chicken, Xenopus
Mol.Wt.: 60kDa; 60kD(Calculated).
Uniprot: P12931
RRID: AB_2834597

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Product Info

Source:
Rabbit
Application:
WB 1:500-1:2000, IHC 1:50-1:200, IF/ICC 1:100-1:500
*The optimal dilutions should be determined by the end user.
*Tips:

WB: For western blot detection of denatured protein samples. IHC: For immunohistochemical detection of paraffin sections (IHC-p) or frozen sections (IHC-f) of tissue samples. IF/ICC: For immunofluorescence detection of cell samples. ELISA(peptide): For ELISA detection of antigenic peptide.

Reactivity:
Human,Mouse,Rat
Prediction:
Pig(100%), Zebrafish(100%), Bovine(100%), Horse(100%), Sheep(100%), Rabbit(100%), Dog(100%), Chicken(100%), Xenopus(100%)
Clonality:
Polyclonal
Specificity:
Phospho-Src (Tyr419) Antibody detects endogenous levels of Src only when phosphorylated at Tyrosine 419.
RRID:
AB_2834597
Cite Format: Affinity Biosciences Cat# AF3162, RRID:AB_2834597.
Conjugate:
Unconjugated.
Purification:
The antibody is from purified rabbit serum by affinity purification via sequential chromatography on phospho-peptide and non-phospho-peptide affinity columns.
Storage:
Rabbit IgG in phosphate buffered saline , pH 7.4, 150mM NaCl, 0.02% sodium azide and 50% glycerol. Store at -20 °C. Stable for 12 months from date of receipt.
Alias:

Fold/Unfold

ASV; Avian sarcoma virus; c SRC; CDNA FLJ14219 fis clone NT2RP3003800 highly similar to Rattus norvegicus tyrosine protein kinase pp60 c src mRNA; cSrc; EC 2.7.10.2; Neuronal CSRC tyrosine specific protein kinase; Neuronal SRC; Oncogene SRC; OTTHUMP00000174476; OTTHUMP00000174477; p60 Src; p60-Src; p60Src; pp60c src; pp60c-src; pp60csrc; Proto oncogene tyrosine protein kinase Src; Proto-oncogene c-Src; Proto-oncogene tyrosine-protein kinase Src; Protooncogene SRC; Protooncogene SRC Rous sarcoma; Src; SRC Oncogene; SRC proto oncogene non receptor tyrosine kinase; SRC_HUMAN; SRC1; Tyrosine kinase pp60c src; Tyrosine protein kinase SRC 1; Tyrosine protein kinase SRC1; v src avian sarcoma (Schmidt Ruppin A2) viral oncogene homolog; V src sarcoma (Schmidt Ruppin A 2) viral oncogene homolog (avian); v src sarcoma (Schmidt Ruppin A 2) viral oncogene homolog avian;

Immunogens

Immunogen:

A synthesized peptide derived from human Src around the phosphorylation site of Tyr419.

Uniprot:
Gene(ID):
Expression:
P12931 SRC_HUMAN:

Expressed ubiquitously. Platelets, neurons and osteoclasts express 5-fold to 200-fold higher levels than most other tissues.

Description:
This gene is highly similar to the v-src gene of Rous sarcoma virus. This proto-oncogene may play a role in the regulation of embryonic development and cell growth. The protein encoded by this gene is a tyrosine-protein kinase whose activity can be inhibited by phosphorylation by c-SRC kinase.
Sequence:
MGSNKSKPKDASQRRRSLEPAENVHGAGGGAFPASQTPSKPASADGHRGPSAAFAPAAAEPKLFGGFNSSDTVTSPQRAGPLAGGVTTFVALYDYESRTETDLSFKKGERLQIVNNTEGDWWLAHSLSTGQTGYIPSNYVAPSDSIQAEEWYFGKITRRESERLLLNAENPRGTFLVRESETTKGAYCLSVSDFDNAKGLNVKHYKIRKLDSGGFYITSRTQFNSLQQLVAYYSKHADGLCHRLTTVCPTSKPQTQGLAKDAWEIPRESLRLEVKLGQGCFGEVWMGTWNGTTRVAIKTLKPGTMSPEAFLQEAQVMKKLRHEKLVQLYAVVSEEPIYIVTEYMSKGSLLDFLKGETGKYLRLPQLVDMAAQIASGMAYVERMNYVHRDLRAANILVGENLVCKVADFGLARLIEDNEYTARQGAKFPIKWTAPEAALYGRFTIKSDVWSFGILLTELTTKGRVPYPGMVNREVLDQVERGYRMPCPPECPESLHDLMCQCWRKEPEERPTFEYLQAFLEDYFTSTEPQYQPGENL

Predictions

Predictions:

Score>80(red) has high confidence and is suggested to be used for WB detection. *The prediction model is mainly based on the alignment of immunogen sequences, the results are for reference only, not as the basis of quality assurance.

Species
Results
Score
Pig
100
Horse
100
Bovine
100
Sheep
100
Dog
100
Xenopus
100
Zebrafish
100
Chicken
100
Rabbit
100
Model Confidence:
High(score>80) Medium(80>score>50) Low(score<50) No confidence

Research Backgrounds

Function:

Non-receptor protein tyrosine kinase which is activated following engagement of many different classes of cellular receptors including immune response receptors, integrins and other adhesion receptors, receptor protein tyrosine kinases, G protein-coupled receptors as well as cytokine receptors. Participates in signaling pathways that control a diverse spectrum of biological activities including gene transcription, immune response, cell adhesion, cell cycle progression, apoptosis, migration, and transformation. Due to functional redundancy between members of the SRC kinase family, identification of the specific role of each SRC kinase is very difficult. SRC appears to be one of the primary kinases activated following engagement of receptors and plays a role in the activation of other protein tyrosine kinase (PTK) families. Receptor clustering or dimerization leads to recruitment of SRC to the receptor complexes where it phosphorylates the tyrosine residues within the receptor cytoplasmic domains. Plays an important role in the regulation of cytoskeletal organization through phosphorylation of specific substrates such as AFAP1. Phosphorylation of AFAP1 allows the SRC SH2 domain to bind AFAP1 and to localize to actin filaments. Cytoskeletal reorganization is also controlled 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 a number of focal adhesion proteins, including PTK2/FAK1 and paxillin (PXN). In addition to phosphorylating focal adhesion proteins, SRC is also active at the sites of cell-cell contact 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 regulation of pre-mRNA-processing and phosphorylates RNA-binding proteins such as KHDRBS1 (Probable). Also plays a role in PDGF-mediated tyrosine phosphorylation of both STAT1 and STAT3, leading to increased DNA binding activity of these transcription factors (By similarity). Involved in the RAS pathway through phosphorylation of RASA1 and RASGRF1. Plays a role in EGF-mediated calcium-activated chloride channel activation. Required for epidermal growth factor receptor (EGFR) internalization through phosphorylation of clathrin heavy chain (CLTC and CLTCL1) at 'Tyr-1477'. Involved in beta-arrestin (ARRB1 and ARRB2) desensitization through phosphorylation and activation of GRK2, leading to beta-arrestin phosphorylation and internalization. Has a critical role in the stimulation of the CDK20/MAPK3 mitogen-activated protein kinase cascade by epidermal growth factor (Probable). Might be involved not only in mediating the transduction of mitogenic signals at the level of the plasma membrane but also in controlling progression through the cell cycle via interaction with regulatory proteins in the nucleus. Plays an important 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. 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. Promotes energy production in osteoclasts by activating mitochondrial cytochrome C oxidase. Phosphorylates DDR2 on tyrosine residues, thereby promoting its subsequent autophosphorylation. Phosphorylates RUNX3 and COX2 on tyrosine residues, TNK2 on 'Tyr-284' and CBL on 'Tyr-731'. Enhances DDX58/RIG-I-elicited antiviral signaling. Phosphorylates PDPK1 at 'Tyr-9', 'Tyr-373' and 'Tyr-376'. Phosphorylates BCAR1 at 'Tyr-128'. Phosphorylates CBLC at multiple tyrosine residues, phosphorylation at 'Tyr-341' activates CBLC E3 activity. Involved in anchorage-independent cell growth. Required for podosome formation (By similarity).

PTMs:

Myristoylated at Gly-2, and this is essential for targeting to membranes.

Dephosphorylated at Tyr-530 by PTPRJ (By similarity). Phosphorylated on Tyr-530 by c-Src kinase (CSK). The phosphorylated form is termed pp60c-src. Dephosphorylated by PTPRJ at Tyr-419. Normally maintained in an inactive conformation with the SH2 domain engaged with Tyr-530, the SH3 domain engaged with the SH2-kinase linker, and Tyr-419 dephosphorylated. Dephosphorylation of Tyr-530 as a result of protein tyrosine phosphatase (PTP) action disrupts the intramolecular interaction between the SH2 domain and Tyr-530, Tyr-419 can then become autophosphorylated, resulting in SRC activation. Phosphorylation of Tyr-530 by CSK allows this interaction to reform, resulting in SRC inactivation. CDK5-mediated phosphorylation at Ser-75 targets SRC to ubiquitin-dependent degradation and thus leads to cytoskeletal reorganization. Phosphorylated by PTK2/FAK1; this enhances kinase activity. Phosphorylated by PTK2B/PYK2; this enhances kinase activity.

S-nitrosylation is important for activation of its kinase activity.

Ubiquitinated in response to CDK5-mediated phosphorylation. Ubiquitination mediated by CBLC requires SRC autophosphorylation at Tyr-419 and may lead to lysosomal degradation.

Subcellular Location:

Cell membrane>Lipid-anchor. Mitochondrion inner membrane. Nucleus. Cytoplasm>Cytoskeleton. Cytoplasm>Perinuclear region. Cell junction>Focal adhesion.
Note: Localizes to focal adhesion sites following integrin engagement (PubMed:22801373). Localization to focal adhesion sites requires myristoylation and the SH3 domain (PubMed:7525268). Colocalizes with PDLIM4 at the perinuclear region, but not at focal adhesions (PubMed:19307596).

Extracellular region or secreted Cytosol Plasma membrane Cytoskeleton Lysosome Endosome Peroxisome ER Golgi apparatus Nucleus Mitochondrion Manual annotation Automatic computational assertionSubcellular location
Tissue Specificity:

Expressed ubiquitously. Platelets, neurons and osteoclasts express 5-fold to 200-fold higher levels than most other tissues.

Family&Domains:

The SH2 and SH3 domains are important for the intramolecular and intermolecular interactions that regulate catalytic activity, localization, and substrate recruitment.

Belongs to the protein kinase superfamily. Tyr protein kinase family. SRC subfamily.

Research Fields

· Cellular Processes > Transport and catabolism > Endocytosis.   (View pathway)

· Cellular Processes > Cellular community - eukaryotes > Focal adhesion.   (View pathway)

· Cellular Processes > Cellular community - eukaryotes > Adherens junction.   (View pathway)

· Cellular Processes > Cellular community - eukaryotes > Tight junction.   (View pathway)

· Cellular Processes > Cellular community - eukaryotes > Gap junction.   (View pathway)

· Cellular Processes > Cell motility > Regulation of actin cytoskeleton.   (View pathway)

· Environmental Information Processing > Signal transduction > ErbB signaling pathway.   (View pathway)

· Environmental Information Processing > Signal transduction > Rap1 signaling pathway.   (View pathway)

· Human Diseases > Drug resistance: Antineoplastic > EGFR tyrosine kinase inhibitor resistance.

· Human Diseases > Drug resistance: Antineoplastic > Endocrine resistance.

· Human Diseases > Infectious diseases: Bacterial > Bacterial invasion of epithelial cells.

· Human Diseases > Infectious diseases: Bacterial > Epithelial cell signaling in Helicobacter pylori infection.

· Human Diseases > Infectious diseases: Bacterial > Shigellosis.

· Human Diseases > Infectious diseases: Bacterial > Tuberculosis.

· Human Diseases > Infectious diseases: Viral > Hepatitis B.

· Human Diseases > Cancers: Overview > Viral carcinogenesis.

· Human Diseases > Cancers: Overview > Proteoglycans in cancer.

· Human Diseases > Cancers: Specific types > Bladder cancer.   (View pathway)

· Organismal Systems > Immune system > Chemokine signaling pathway.   (View pathway)

· Organismal Systems > Development > Axon guidance.   (View pathway)

· Organismal Systems > Immune system > Platelet activation.   (View pathway)

· Organismal Systems > Nervous system > GABAergic synapse.

· Organismal Systems > Sensory system > Inflammatory mediator regulation of TRP channels.   (View pathway)

· Organismal Systems > Endocrine system > Estrogen signaling pathway.   (View pathway)

· Organismal Systems > Endocrine system > Prolactin signaling pathway.   (View pathway)

· Organismal Systems > Endocrine system > Thyroid hormone signaling pathway.   (View pathway)

· Organismal Systems > Endocrine system > Oxytocin signaling pathway.

· Organismal Systems > Endocrine system > Relaxin signaling pathway.

References

1). Galectin-3 induces pathogenic immunosuppressive macrophages through interaction with TREM2 in lung cancer. Journal of experimental & clinical cancer research : CR, 2024 (PubMed: 39135069) [IF=11.3]

Application: WB    Species: Mouse    Sample:

Fig. 4 Galectin-3 inhibits TREM2/DAP12 receptor complex to suppress Src/Syk signaling pathway and altered macrophage to an M2-like phenotype. (A, B) 293T cells were transfected with plasmids as shown, and anti-HA (B) or anti-Flag (B) antibodies were employed for exogenous CO-IP experiments. (C-E) 293T cells were transfected with plasmids as shown and anti-HA (C), anti-Flag (D), or anti-Myc (E) antibodies were employed for exogenous CO-IP experiments, respectively. (F, G) After 24 h of LLC CM treatment with the addition of GB1107 (5 µM), and 2 h of stimulation with LPS (1 ng/mL), the phosphorylation levels of Src and Syk in indicated time point were analyzed by western blot (F). The gray values of p-Syk and p-Src protein bands were analyzed by Image J software, and the relative gray values were standardized to the gray values of Syk and Src (G). (H, I) After 24 h of LLC CM treatment with the addition of GB1107 (5 µM), and 2 h of stimulation with LPS (1 ng/mL), the phosphorylation levels of Src and Syk in WT or TREM2 KO BMDMs were analyzed by western blot (H). The gray values of p-Syk and p-Src protein bands were analyzed by Image J image analyses software, and the relative gray values were standardized to the gray values of Syk and Src (I). (J, K) After 24 h of LLC CM treatment with the addition of rm galectin-3 (200 ng/ml), and 2 h of stimulation with LPS (1 ng/mL), the phosphorylation levels of Src and Syk in WT or TREM2 KO BMDMs were analyzed by western blot (J). The gray values of p-Syk and p-Src protein bands were analyzed by Image J image analyses software, and the relative gray values were standardized to the gray values of Syk and Src (K). (L) Phagocytosis assay between WT or TREM2 KO BMDMs which were pretreated with LLC CM supplemented with GB1107 (5 µM) for 24 h and LLC were detected by flow cytometry. (M) Following a 24 h-treatment with LLC CM supplemented with GB1107 (5 µM), the F-actin polarization of RAW264.7 cells blocked with anti-TREM2 antibody was observed by confocal microscopy. Scale bars, 5 μm. Quantitative statistics of F-actin polarization were analyzed by Image J image analyses software. (N-P) After 24 h of LLC CM which was supplemented with GB1107 (5 µM) treatment, the transcription levels of Ccr2(N), M2-like macrophage markers (CD206, Arg1) (O) and M1-like macrophage markers (Nos2, TNFα) (P) in WT or TREM2 KO BMDMs were detected by RT-qPCR assay. Data represent mean ± SD from three experiments. *, P 

2). Src acts as the target of matrine to inhibit the proliferation of cancer cells by regulating phosphorylation signaling pathways. Cell Death & Disease, 2021 (PubMed: 34642304) [IF=8.1]

Application: WB    Species: Human    Sample: HT-29, MCF7, A549, BxPC-3, SKOV3, and HeLa cells

Fig. 4 Matrine was a non-ATP-competitive inhibitor of Src kinase. A The effect of matrine on Src kinase activity in cancer cells, including HT-29, MCF7, A549, BxPC-3, SKOV3, and HeLa, were determined according to the instructions of Src kinase activity detection kit. Results are representative of three experiments (*P 

3). Anti-proliferation and anti-migration effects of Yishen Tongbi decoction in experimental rheumatoid arthritis by suppressing SLC3A2/integrin β3 signaling pathways. Phytomedicine, 2023 (PubMed: 36990010) [IF=6.7]

4). Metastasis suppressor 1 controls osteoblast differentiation and bone homeostasis through regulating Src-Wnt/β-catenin signaling. Cellular and molecular life sciences : CMLS, 2022 (PubMed: 35094173) [IF=6.2]

Application: WB    Species: Mouse    Sample: ST2 cells

Fig. 4 MTSS1 inactivated Src and activated Wnt/β-catenin signaling in stromal progenitor cells. ST2 cells were transfected with MTSS1 expression construct (A, D) or MTSS1 siRNA (B, E) or Src expression construct (F) and their control. The protein levels of MTSS1, phosphorylated, and total Src were detected in the cells 3 days after transfection using Western blotting (A, B). Co-IP was performed and endogenous MTSS1 and LRP6 were detected with Western blotting (C). The protein levels of the major components of Wnt/β-catenin signaling were detected in the cells 3 days after transfection using Western blotting (D–F). Values represent mean ± SD, n = 3. Student’s t test was conducted in (A, D, F). One-way ANOVA was conducted in (B, E) followed by Tukey test for post hoc comparisons. *Significant vs. vector or control siRNA, p 

5). Formononetin Protects LPS-Induced Mastitis Through Suppressing Inflammation and Enhancing Blood-Milk Barrier Integrity via AhR-Induced Src Inactivation. Frontiers in Immunology, 2022 (PubMed: 35185907) [IF=5.7]

Application: WB    Species: Mice    Sample:

Figure 8 FOR increases AhR expression and inhibits the phosphorylation level of Src. The present results are displayed as the mean ± SEM of triple parallel measurements. #(P < 0.01) is significantly different from control group; **(P < 0.01) is significantly different from LPS group.

6). Jinfukang regulates integrin/Src pathway and anoikis mediating circulating lung cancer cells migration. JOURNAL OF ETHNOPHARMACOLOGY, 2021 (PubMed: 33068649) [IF=4.8]

Application: WB    Species: human    Sample: CTC-TJH-01 cells

Fig. 5. |Jinfukang inhibited the cell migration via Integrin/Src axis in lung cancer cells. Cells were incubated with Jinfukang (100 μg/mL), ATN-161 (10 μmol/L) or a combination of both for 24 h. A Western blot assay was used to measure the protein expression levels of Integrin β1, Src, and p-Src in CTC-TJH-01 (A) and H1975 (B)cells. GAPDH was used as an internal standard.

7). Cyclin G2 inhibits oral squamous cell carcinoma growth and metastasis by binding to IGFBP3 and regulating the FAK-SRC-STAT signaling pathway. Frontiers in Oncology, 2020 (PubMed: 33240810) [IF=3.5]

Application: WB    Species: Human    Sample: Oral squamous cell carcinoma (OSCC)

FIGURE 5 | Cyclin G2 inhibits the FAK-SRC-STAT pathway in vitro and in vivo. (A) The protein expressions along with quantification of FAK, P-FAK, SRC, P-SRC, STAT3, P-STAT3, Bcl-2, c-Myc, and MMP9 in the FAK-SRC-STAT pathway. (B–D) The relative mRNA expressions of Bcl-2, c-Myc and MMP9. (E) Analysis of MMP9 secretion and its quantification in SCC-9 cells. (F, G) Immunohistochemical staining and analysis of the expression of cyclin G2, p-FAK, p-SRC and p-STAT3. Scale bar = 100 µm. *p < 0.05, **p < 0.01, ***p < 0.001 vs. vector.

8). Ethoxy-erianin phosphate and afatinib synergistically inhibit liver tumor growth and angiogenesis via regulating VEGF and EGFR signaling pathways. Toxicology and Applied Pharmacology, 2022 (PubMed: 35143806) [IF=3.3]

9). Inorganic arsenic exposure promotes malignant progression by HDAC6-mediated down-regulation of HTRA1. Journal of Applied Toxicology, 2023 (PubMed: 36861143) [IF=2.7]

Application: WB    Species: Human    Sample: Caco-2 cells

FIGURE 3 Molecular profiling for iAs-induced changes in Caco-2 cells. (A) Protein levels of claudin 3 and cofilin were determined by Western blot analysis. (B) Transcript levels of IL-6, TNF-α, and MMP13 were determined by RT-qPCR, with GAPDH as an internal control. (C) The levels of EGF in the culture supernatant were examined by ELISA. (D) A marked increase in the phosphorylated levels of EGFR, Src, AKT, and p38 was shown by Western blot analysis. RT-qPCR (E) and Western blot analysis (F) results showed the down-regulation of HTRA1 expression caused by chronic iAs exposure. Data were representatives of at least three independent experiments, shown as mean ± SD. The significance threshold for Student's t-test:

10). Overexpression of Laminin 5γ2 Chain Correlates with Tumor Cell Proliferation, Invasion, and Poor Prognosis in Laryngeal Squamous Cell Carcinoma. Journal of Oncology, 2022 (PubMed: 36284634)

Application: WB    Species: Human    Sample: AMC-HN-8 cells

Figure 6 LAMC2 activated the integrin β1/FAK/Src/AKT signaling pathway in TU177 and AMC-HN-8 cells. (a) LAMC2 overexpression promotes integrin β1/FAK/Src/AKT protein expression in TU177 cells. (b) LAMC2 knockdown inhibited integrin β1/FAK/Src/AKT protein expression in AMC-HN-8 cells. Data were expressed as mean ± SD, n = 3. Compared to the vector group, #P < 0.05, ##P < 0.01. Compared to the shNC group, ∗P < 0.05, ∗∗P < 0.01.

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