Custom phosphorylation antibody
Post-translational modification plays an important role in many key regulatory mechanisms of life phenomena. Among them, protein phosphorylation is one of the most common post-translational modifications. It takes part in almost all cell activities in the body, including signal transduction, cell cycle, proliferation and metabolism, growth and development, cancer mechanism and many other biological problems Close relationship.Because phosphorylated proteins can play a complex role, it is important to accurately and accurately quantify phosphorylated proteins at the cellular level for deciphering and understanding cell functions.
Protein phosphorylation refers to the process of transferring the phosphorylation of ATP molecules to the specific sites of protein molecules (SER, THR, Tyr residues, etc.) through protein phosphokinase.Some studies have shown that the interaction between proteins can be regulated by phosphorylation of multiple sites.The abnormal up or down regulation of protein phosphorylation is closely related to the occurrence of many diseases, such as cancer, diabetes, metabolic syndrome and Alzheimer's disease.Therefore, it is necessary to study the characteristics of protein phosphorylation in various biological and abiotic stress fields.
Phosphorylated antibodies are mainly prepared for phosphorylation sites, which can specifically recognize the phosphorylated amino acid sites, detect the phosphorylation level of proteins, and carry out qualitative and quantitative analysis of phosphorylated proteins.Phosphorylated antibody can be used to detect the change of phosphorylation level of the protein when the cell is stimulated, focusing on the understanding of an active state of the protein, which is widely used in immunology, biochemistry, oncology, proteomics, especially in the study of cell signal transduction, withering, cancer and other life sciences.
Trigoats provides customized services for phosphorylated antibodies including: Phosphorylated peptide design and synthesis, animal coupling immunity, monoclonal screening and library building, antibody production and purification, antibody quality verification and delivery.The final delivered antibody will be confirmed by the double detection of phosphorylated peptide and non phosphorylated peptide to ensure the quality. The phosphorylated antibody can specifically recognize the phosphorylated amino acid sites.Phosphorylated antibody can be used to distinguish the existing forms of protein phosphorylation and non phosphorylation, and can be used for qualitative and quantitative analysis of phosphorylated protein. Trigoats' professional antibody R & D team has rich experience in analysis, design, synthesis of phosphorylated polypeptide and preparation technology of phosphorylated antibody.We provide phosphorylated polyclonal antibody service and phosphorylated monoclonal antibody service.n the customized service of phosphorylated multi antibody, the serum collected after animal immunization was purified by phosphorylated peptide affinity chromatography, and then the preliminary purified antibody was absorbed by non phosphorylated peptide affinity chromatography. The final delivered phosphorylated antibody was confirmed by double detection of BSA cross-linked phosphorylated peptide and non phosphorylated peptide, thus ensuring the delivery quality of phosphorylated antibody.
Preparation of phosphorylated polyclonal antibody (rabbit)
Service phase |
service content |
Deliver products |
Stage 1
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Design and synthesis of phosphorylated polypeptide 10 mg phosphorylated polypeptide and 5 mg non phosphorylated polypeptide
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• 2-3 mg peptides (nonphosphorylated and phosphorylated)
• 1-2 ml pre immune serum
• 2-5mg phosphorylated antibody
• ELISA results
• Project Report
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Stage 2
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Peptide coupling Conjugation of synthetic peptide and carrier protein (KLH, BSA)
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Stage 3
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preparation of antiserum Immunize rabbits, collect blood before immunizing, immunize four times and collect serum
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Stage 4
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Small scale purification Collection and detection of serum samples
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Stage 5
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Antigen affinity purification Preparation of phosphorylated affinity purification column and removal of cross reaction by non phosphorylated peptide affinity column
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Stage 6
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Testing Titer of ELISA
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Preparation of phosphorylated monoclonal antibody (mouse)mei
Service phase
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service content
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Deliver products
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Stage 1
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Design and synthesis of phosphorylated polypeptide 10 mg phosphorylated polypeptide and 5 mg non phosphorylated polypeptid |
• 5mg modified peptide, 3mg unmodified peptide and antibody products
• 50ul pre immune serum
• 2-5 hybridoma cells
• ELISA results of 3-8mg purified antibody (2 hybridomas)
• Subtype identification results
• Full project report
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Stage 2 | Peptide coupling c |
Stage 3 | Animal immunity and detection Immunized mice and detected by ELISA with phosphorylated and non phosphorylated polypeptides |
Stage 4 | The best immune response shown by cell fusion phosphorylated polypeptides: cell fusion of spleen cells and myeloma cells in mice |
Stage 5 | Proliferation and screening The positive clones were detected by ELISA and |
Stage 6 | Ascites preparation and purification ascites preparation, protein G purification |
Stage 7 | Detection ELISA detection titer |