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Die folgenden MAPmates™ sollten nicht zusammen analysiert werden: -MAPmates™, die einen unterschiedlichen Assaypuffer erfordern. -Phosphospezifische und MAPmate™ Gesamtkombinationen wie Gesamt-GSK3β und Gesamt-GSK3β (Ser 9). -PanTyr und locusspezifische MAPmates™, z.B. Phospho-EGF-Rezeptor und Phospho-STAT1 (Tyr701). -Mehr als 1 Phospho-MAPmate™ für ein einziges Target (Akt, STAT3). -GAPDH und β-Tubulin können nicht mit Kits oder MAPmates™, die panTyr enthalten, analysiert werden.
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48-602MAG
Buffer Detection Kit for Magnetic Beads
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Anti-Phosphotyrosine (Fab fragment), clone 4G10-G6 Cat. No. MABS2179, is a recombinant monoclonal antibody that detects phosphotyrosines in protein and is tested for use in Immunocytochemistry, Immunohistochemistry, and Western Blotting.
More>>Anti-Phosphotyrosine (Fab fragment), clone 4G10-G6 Cat. No. MABS2179, is a recombinant monoclonal antibody that detects phosphotyrosines in protein and is tested for use in Immunocytochemistry, Immunohistochemistry, and Western Blotting. Less<<
Phosphorylation on tyrosine residues is one of the major signal transduction mechanism that regulates intercellular and intracellular signaling in multicellular organisms. Dysregulation of protein tyrosine phosphorylation leads to several diseases, including cancer, diabetes, and several neurodegenerative disorders. Tyrosine residues in proteins can be phosphorylated by protein tyrosine kinases resulting in the formation of phosphotyrosine. Tyrosine phosphorylation is one of the key steps in signal transduction and regulation of enzymatic activity. Anti-phosphotyrosine antibodies are commonly used in Western blotting application after the targeted proteins have been immunoprecipitated to measure the tyrosyl phosphorylation of the proteins. They can also be used directly in cell lysates to examine the overall change of tyrosine phosphorylation level in response to various hormones, cytokines, and other treatments. Clone 4G10® is one of the best monoclonal antibodies for the detection tyrosine phosphorylation. This clone provides high sensitivity and can detect tyrosine phosphorylation of multiple proteins. It interacts only with the phosphotyrosine (pY) residue and not with any neighboring ones. Clone 4G10-G6 is recombinantly expressed using E. coli in a Fab format and offers about 7-fold improved signal over parental 4G10. It allows for the detection of difficult to detect bands, particularly in the range of 25 to 55 kDa when used with anti-His Tag secondary antibody. (Ref.: Mou, Y., et al. (2018). J. Am. Chem. Soc. 140, 16615-16624.
References
Product Information
Format
Purified
Presentation
Recombinant monoclonal antibody Fab in buffer containing 0.1 M Tris-Glycine (pH 7.4), 150 mM NaCl with 0.05% sodium azide.
Anti-Phosphotyrosine (Fab fragment), clone 4G10-G6 Cat. No. MABS2179, is a recombinant monoclonal antibody that detects phosphotyrosines in protein and is tested for use in Immunocytochemistry, Immunohistochemistry, and Western Blotting.
Key Applications
Western Blotting
Immunocytochemistry
Immunohistochemistry
Application Notes
Immunocytochemistry Analysis: A representative lot detected Phosphotyrosine in HeLa cells treated with 0.1 mM pervanadate. (Mou, Y., et al. (2018). J. Am. Chem. Soc. 140(48); 16615-16624).
Immunohistochemistry (Paraffin) Analysis (IHC(P)): 3 µg/mL of this antibody detected Phosphotyrosine in human breast cancer tissue sections using an anti-His Tag secondary antibody and HRP-DAB detection system.
Note: Actual optimal working dilutions must be determined by end user as specimens, and experimental conditions may vary with the end user
Biological Information
Immunogen
Recombinant antibody from Fabs constructed in a dual expression vector for Light chain and His-tagged heavy chain of clone 4G10 and expressed in C43(DE3) strain of E. coli.
Epitope
Phosphotyrosine
Clone
4G10-G6
Concentration
1 mg/mL. Please refer to guidance on suggested starting dilutions and/or titers per application and sample type.
Host
E. coli
Specificity
Clone 4G10-G6 is a his tagged recombinant monoclonal antibody that detects phosphotyrosine in proteins from all species. To be used with anti-His Tag secondary antibody.
Species Reactivity
All
Species Reactivity Note
Predicted to react with all species.
Antibody Type
Monoclonal Antibody
Modifications
Phosphorylation
Purification Method
Immobilized Metal Affinity Chromatography (IMAC)
Physicochemical Information
Dimensions
Materials Information
Toxicological Information
Safety Information according to GHS
Safety Information
Product Usage Statements
Quality Assurance
Evaluated by Western Blotting in lysate from Jurkat cells treated with pervanadate.
Western Blotting Analysis: 1 µg/mL of this antibody detected Phosphotyrosine in lysate of Jurkat cells treated with 0.1 mM pervanadate.
Usage Statement
Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.
Storage and Shipping Information
Storage Conditions
Stable for 1 year at +2°C to +8°C from date of receipt.
Phosphotyrosine (pY) is one of the most highly studied posttranslational modifications that is responsible for tightly regulating many signaling pathways in eukaryotes. Pan-specific pY antibodies have emerged as powerful tools for understanding the role of these modifications. Nevertheless, structures have not been reported for pan-specific pY antibodies, greatly impeding the further development of tools for integrating this ubiquitous posttranslational modification using structure-guided designs. Here, we present the first crystal structures of two widely utilized pan-specific pY antibodies, PY20 and 4G10. The two antibodies, although developed independently from animal immunizations, have surprisingly similar modes of recognition of the phosphate group, implicating a generic binding structure among pan-specific pY antibodies. Sequence alignments revealed that many pY binding residues are predominant in the mouse V germline genes, which consequently led to the convergent antibodies. On the basis of the convergent structure, we designed a phage display library by lengthening the CDR-L3 loop with the aid of computational modeling. Panning with this library resulted in a series of 4G10 variants with 4 to 11-fold improvements in pY binding affinities. The crystal structure of one improved variant showed remarkable superposition to the computational model, where the lengthened CDR-L3 loop creates an additional hydrogen bond indirectly bound to the phosphate group via a water molecule. The engineered variants exhibited superior performance in Western blot and immunofluorescence.
