Immunofluorescence analysis of SRC [pY529] was done on 70% confluent log phase A549 cells. The cells were fixed with 4% paraformaldehyde for 15 minutes, permeabilized with 0.25% Triton™ X-100 for 10 minutes, and blocked with 5% BSA for 1 hour at room temperature. The cells were labeled with SRC [pY529] Rabbit Polyclonal Antibody (44662G) at 1ug/mL in 1% BSA and incubated for 3 hours at room temperature and then labeled with Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A27034) at a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (S36938). F-actin (Panel c: red) was stained with Alexa Fluor® 555 Rhodamine Phalloidin (Product # R415, 1:300). Panel d is a merged image showing Nuclear localization. Panel e is a no primary antibody control. The images were captured at 60X magnification.
|Tested species reactivity||Chicken, Human, Mouse, Rat|
|Published species reactivity||Rat, Bovine, Human, Mouse|
|Host / Isotype||Rabbit / IgG|
|Immunogen||The antiserum was produced against a chemically synthesized phosphopeptide derived from the region of human Src that contains tyrosine 529 (tyrosine 530 including the initiating methionine). The sequence is conserved in mouse, rat, chicken and frog.|
|Purification||Antigen affinity chromatography|
|Storage buffer||Dulbecco's PBS, pH 7.3, with 50% glycerol, 1mg/ml BSA|
|Contains||0.05% sodium azide|
|Tested Applications||Dilution *|
|Flow Cytometry (Flow)||3-5 µg/million cells|
|Immunocytochemistry (ICC)||1 µg/ml|
|Immunofluorescence (IF)||1 µg/ml|
|Immunohistochemistry (Paraffin) (IHC (P))||1:50-1:500|
|Western Blot (WB)||1-2 µg/mL|
* Suggested working dilutions are given as a guide only. It is recommended that the user titrate the product for use in their own experiment using appropriate negative and positive controls.
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. Mutations in this gene could be involved in the malignant progression of colon cancer. Two transcript variants encoding the same protein have been found for this gene. Transcript Variant: This variant (1) represents the longer transcript. Both isoforms 1 and 2 encode the same protein.
Altered expression of proteins upon cell treatment demonstrates antibody specificity.Western blot of Phospho-SRC (Tyr529) using Anti - SRC [pY529] Rabbit polyclonal Antibody (Product # 44-662G), shows increase in expression of SRC [pY529] upon treatment with PDGF for a synchronised A549 and SHSY5Y cell population.Also, increase in expression of Phospho-SRC (Tyr529) is observed upon treatment EGF for a HT-29 synchronised cell population. Cell Treatment validation info.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
Fyn is an intermediate kinase that BDNF utilizes to promote oligodendrocyte myelination.
44-662G was used in western blot to study how BDNF activation and phosphorylation of Fyn regulate oligodendrocyte myelination.
|Peckham H,Giuffrida L,Wood R,Gonsalvez D,Ferner A,Kilpatrick TJ,Murray SS,Xiao J||Glia (64:255)||2016|
Phosphatidylinositol phosphate 5-kinase I¿i2 in association with Src controls anchorage-independent growth of tumor cells.
44-662G was used in immunoprecipitation and western blot to study how tumor cells sustain oncogenic signals in the absence of cell matrix interactions.
|Thapa N,Choi S,Hedman A,Tan X,Anderson RA||The Journal of biological chemistry (288:34707)||2013|
||Analyzing FAK and Pyk2 in early integrin signaling events.||Bernard-Trifilo JA,Lim ST,Hou S,Schlaepfer DD,Ilic D||Current protocols in cell biology (Chapter 14:null)||2006|
||A mechanosensory complex that mediates the endothelial cell response to fluid shear stress.||Tzima E,Irani-Tehrani M,Kiosses WB,Dejana E,Schultz DA,Engelhardt B,Cao G,DeLisser H,Schwartz MA||Nature (437:426)||2005|
|Mouse||Not Cited||An osteoclastic protein-tyrosine phosphatase regulates the ß3-integrin, syk, and shp1 signaling through respective src-dependent phosphorylation in osteoclasts.||Lau KH,Stiffel V,Amoui M||American journal of physiology. Cell physiology (302:C1676)||2012|
|Mouse||Not Cited||Analyzing FAK and Pyk2 in early integrin signaling events.||Bernard-Trifilo JA,Lim ST,Hou S,Schlaepfer DD,Ilic D||Current protocols in cell biology (Chapter 14:null)||2006|
|Bovine||Not Cited||A mechanosensory complex that mediates the endothelial cell response to fluid shear stress.||Tzima E,Irani-Tehrani M,Kiosses WB,Dejana E,Schultz DA,Engelhardt B,Cao G,DeLisser H,Schwartz MA||Nature (437:426)||2005|
|Mouse||Not Cited||Reelin activates SRC family tyrosine kinases in neurons.||Bock HH,Herz J||Current biology : CB (13:18)||2003|