The Gold Standard in Anti-Phosphotyrosine Antibodies
4G10 Anti-Phosphotyrosine was originally developed in 1989 by one of the world’s leading cell signaling researchers, Dr. Brian Druker. The development of 4G10 Anti-Phosphotyrosine was monumental for researchers as it facilitated the study of tyrosine kinase activity by simple blotting procedures, without the need for radioactivity. Over the last 15 years, 4G10 has been validated in virtually every application and tyrosine target by thousands of scientific and medical researchers. 4G10 Anti-Phosphotyrosine is now an important tool for research in areas as diverse as serine/ threonine phospho-proteins (including p38, Akt and ERK1/2), G-proteins and neurological targets.
Cellular Signal Transduction through Tyrosine Phosphorylation Protein phosphorylation is often necessary for the initiation of biological processes such as cell growth, proliferation, and ubiquitin-mediated degradation. Tyrosine phosphorylation, in particular, often mediates receptor signaling events at the membrane by initiating protein-protein interactions in response to growth factors, hormones and cytokines. Research tools that detect tyrosine phosphorylation are central in understanding the significance of cellular signaling in many biological processes and disease origination.
4G10 Anti-Phosphotyrosine Conjugates are Available to Facilitate and Accelerate Your Research
Both native and recombinant 4G10 antibodies are now offered in conjugated formats, including agarose, biotin, FITC and HRP, to accelerate your research. With the Upstate® Grand of 4G10 antibody, spend more time on primary research and less time optimizing experimental conditions. Not only will these conjugates save you valuable time in the laboratory, they can also save you money by eliminating the costs of additional antibodies. Contact Millipore’s Technical Support group to see if there is a 4G10 conjugate suitable for your application.
4G10 Anti-Phosphotyrosine is Validated in Nearly Every Application
4G10 Anti-Phosphotyrosine has been characterized in each of the applications listed below and is proven not to cross-react with phospho-serine or phospho-threonine peptides or proteins.
In addition to its unparalleled characterization and applicability, 4G10 Anti-Phosphotyrosine is well known as one of the most sensitive phosphotyrosine antibodies, detecting at least twice as many phosphoproteins in a side-by-side comparison to PY20 and PT-66 by independent investigators. This sensitivity is also observed in other applications, including fluorescence polarization, where Upstate Grand of and recombinant upstate 4G10 antibodies has been shown to be more sensitive than P-Tyr-100.
4G10 Antibodies Detects More Than 30 Tyrosine Kinases
Both native and recombinant 4G10 antibodies have been proven to detect more than 30 phosphorylated tyrosine kinases. Whether you are studying the role that insulin receptors play in diabetes or constitutively active Abl in chronic myelogenous leukemia, 4G10 products are what you need.
| Abl | EphB2 | Insulin R | Rse |
| Arg | EphB4 | Lck | Syk |
| Axl | Fes/Fps | Lyn | Src |
| Blk | FGFR3 | MAPK2 | Tie2 |
| Bmx | Flt3 | Met | TrkB |
| BTK | Fms | PDGFRa | Yes |
| Csk | Fyn | PDGFRß | ZAP-70 |
| EGFR | IGF R | Ros |  |
Tyrosine phosphorylation often mediates receptor signaling events at the membrane by initiating protein-protein interactions in response to growth factors, hormones, and cytokines.
Binding of anti-phosphotyrosine antibodies to a fluorescently labeled phosphotyrosine tracer peptide.
Tyrosine Kinase FP Assay Enzyme Titrations
Each kinase was serially diluted in assay buffer over the concentration range shown. Kinase activity was measured at 25°C in a 384 well plate and stopped by the addition of EDTA, followed by the addition of a common detection buffer containing 4G10 and fluorescein-labelled phosphopeptide (tracer). Activity was measured by fluorescence polarization, a no enzyme control (i.e., 4G10 and tracer, maximum signal) is shown as an orange circle. Increasing kinase activity resulted in a decrease in fluorescence polarization shown as green circles.
4G10 IHC Stain IHC: Paraffin embedded poorly-differentiated adenocarcinoma, stained with Anti-Phosphotyrosine, clone 4G10 (Millipore cat. no. 05-321) at 10 µg/mL. |
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