Product nameAnti-Src (phospho Y529) antibody [Y232]
See all Src primary antibodies
DescriptionRabbit monoclonal [Y232] to Src (phospho Y529)
ab32078 will detect Src phosphorylation on Tyrosine 529 of both isoforms. The antibody immunogen shares 93% homology with Fyn and Yes and 85% homology with Fgr. Therefore, it is likely that the antibody will cross-react with these proteins. However, this is just based on BLAST results and no experiments were performed. The sequence numbering is based off the mature form of the protein without the initiator methionine.
Tested applicationsSuitable for: IP, WB, Dot blot, ICC/IFmore details
Unsuitable for: Flow Cyt or IHC
Species reactivityReacts with: Mouse, Rat, Human
Synthetic peptide within Human Src (phospho Y529). The exact sequence is proprietary. The sequence numbering is based off the mature form of the protein without the initiator methionine.
(Peptide available as
- WB: HeLa and A431 cell lysates ICC/IF: HeLa and A431 cells
Storage instructionsShipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles.
Storage bufferpH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 49% PBS, 50% Glycerol, 0.05% BSA
Concentration information loading...
Immunizing Peptide (Blocking)
Our Abpromise guarantee covers the use of ab32078 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|WB||1/5000 - 1/10000. Predicted molecular weight: 60 kDa.Can be blocked with Human Src (phospho Y529) peptide (ab179556).|
|ICC/IF||1/50 - 1/500.|
FunctionNon-receptor protein tyrosine kinase that plays pivotal roles in numerous cellular processes such as proliferation, migration, and transformation. In concert with PTK2B, plays an important role in osteoclastic bone resorption. Both the formation of a SRC-PTK2B complex, and SRC kinase activity are necessary for this function. Once it is recruited to the activated integrins, by PTK2B, it phosphorylates 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 RUNX3 and COX2 on tyrosine residues, TNK2 on 'Tyr-284' and CBL on 'Tyr-731'. Enhances DDX58/RIG-I-elicited antiviral signaling.
Sequence similaritiesBelongs to the protein kinase superfamily. Tyr protein kinase family. SRC subfamily.
Contains 1 protein kinase domain.
Contains 1 SH2 domain.
Contains 1 SH3 domain.
modificationsDephosphorylated 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.
S-nitrosylation is important for activation of its kinase activity.
Cellular localizationCell membrane. Mitochondrion inner membrane.
- Information by UniProt
FormThis protein is known to be similar in amino acid sequence to HCK (P08631), LCK (P06239), FYN (P06241), YES1 (P07947), and LYN (P07948). Therefore, cross-reactivity with these homologous proteins may be observed. We would be happy to provide immunogen alignment information upon request.
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WB analysis. Lane 1:Untreated HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysates 15ug and Lane 2:HeLa (Human epithelial cell line from cervix adenocarcinoma) treated with H2O2 at 10mM for 1 h. whole cell lysates 15ug and Lane 3:HeLa (Human epithelial cell line from cervix adenocarcinoma) treated with H2O2 at 10mM for 1 h. whole cell lysates 15ug. Then the membrane was incubated with phosphatase. Primary ab used at 1:30,000 dilution. 5% NFDM/TBST was used as Blocking buffer and Diluting buffer. Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) was used as a Secondary ab at 1:20,000 dilution. The Exposure time was 30 seconds.
Immunocytochemistry/Immunofluorescence analysis of HeLa (Human epithelial cell line from cervix adenocarcinoma) cells labelling Src (phospho Y529) with ab32078 at 1/500. Cells were fixed with 4% Paraformaldehyde and permeabilized with 0.1% Triton X-100. ab150077, Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/1000) was used as the secondary antibody. Cells were counter-stained with ab195889 Anti-Alpha Tubulin antibody [DM1A] (1/200) - Microtubule Marker (Alexa Fluor® 594). DAPI (blue) was used as a nuclear counterstain.
The green staining was increased in the H2O2 (10mM, 1 hour) treated HeLa cells when compared with HeLa cells without treatment. After LP treatment, the green signaling was obviously decreased.
For the pan antibody, there was no great difference after H2O2 (10 mM, 1 hour) or EGF (100 ng/mL, 10 minutes) + LP treatment.
The data showed mostly Cytoplasm and Membran staining for Ab32078.
Dot Blot analysis of Lane 1: Src (pY529) phospho peptide and Lane 2: Src non-phospho peptide labeling Src (phospho Y529) with ab32078 at 1/1000 dilution. 5% NFDM/TBST was used as the diluting and blocking buffer. ab97051 Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated was used as the secondary antibody at 1/100000 dilution. Exposure time: 3 minutes.
All lanes : Anti-Src (phospho Y529) antibody [Y232] (ab32078) at 1/10000 dilution
Lane 1 : Untreated A431 cells
Lane 2 : EGF treated A431 cells
Lane 3 : EGF and alkaline phosphatase treated A431 cells
Predicted band size: 60 kDa
Observed band size: 55 kDa why is the actual band size different from the predicted?
Dot Blot analysis on immunogen phospho peptide (A) and non phospho peptide (B) using 1/2000 ab32078.
This product has been referenced in:
- Niu F et al. Cocaine-induced release of CXCL10 from pericytes regulates monocyte transmigration into the CNS. J Cell Biol 218:700-721 (2019). Read more (PubMed: 30626719) »
- Zhou Y et al. CD59 is a potential biomarker of esophageal squamous cell carcinoma radioresistance by affecting DNA repair. Cell Death Dis 9:887 (2018). Read more (PubMed: 30166523) »