Phosphorylated Monoclonal & Polyclonal Antibody Service

KMD Bioscience has extensive experience in producing customized phosphorylated monoclonal and polyclonal antibodies. We can provide antibody services for specific phosphorylation sites as customers’ request. In addition, we provide comprehensive phosphorylation antibody validation services, including phosphorylation and non-phosphorylation antigen cross-reactivity, Western Blot and immunohistochemistry.

 

Phosphorylated Antibody Production Services:

 

Protein phosphorylation is a significant post-translational modification (PTM) in cells, mainly involved in regulating protein biological activity, including activation or inhibition of protein activity, thereby further regulating the cellular signal pathway. In eukaryotes, the phosphorylation sites of proteins are serine, threonine and tyrosine; in prokaryotes, the phosphorylation site is mainly histidine. KMD Bioscience provides customers with a variety of protocols for phosphorylated antibody production, including traditional antibody development, such as murine monoclonal hybridoma development techniques and polyclonal antibody development techniques, as well as recombinant antibody development techniques.

Phosphorylated antibody development services based on traditional antibody production techniques:

Analysis of phosphorylation sites; design and synthesis of phosphorylated and non-phosphorylated polypeptide antigens; animal immunization and serum titer evaluation; antibody purification and pairing services.

 

Phosphorylated Antibody Development Service Protocol: 

 

Procedure

Description

Deliverable

Antigen Design & Synthesis

* Customer provides antigen (polypeptide, protein) with purity >90%

* KMD Bioscience designs and synthesizes phosphorylated and non-phosphorylated antigens (optional)

* MS and HPLC reports of phosphorylated and non-phosphorylated antigens

* 2-5 mg phosphorylated polypeptide, purity >90%

* 2-5 mg non-phosphorylated polypeptide, purity >90%

Monoclonal Abs Development

* Immunize 5 Balb/c mice and serum titer testing by ELISA

* Take spleen cells for cell fusion (sp2/0 or NS0 tumor cells)

* Cell cloning and subcloning, antibody containing cell culture supernatant ELISA titer & cross reactivity validation

* 2-5 antibody hybridoma cell lines

* 2-5 mg purified antibody

* ELISA report

* No less than one monoclonal antibody,1000-fold difference in recognization efficiency between phosphorylated and non-phosphorylated antigens

* Western Blot validation report (optional)

Polyclonal Abs Development

* Immunization of 2-3 New Zealand white rabbits (using a unique rabbit-specific immune adjuvant, producing high antibody titers in 28 days), ELISA test serum titer

* Serum collection and ELISA titer detection

* Antigen affinity purification (recommended) or protein A purification

* Validation of antibody dilution ratio (customer samples are required)

* 1-2ml pre-immune serum

* 1-5 mg purified antibody

* ELISA report

* Antibody of 1000-fold difference in recognization efficiency between phosphorylated and non-phosphorylated antigens

* Western Blot validation report (optional)

 

Service Highlights:

 

--Fully traceable document support system, GMP/GLP level

--High antibody titer: Guaranteed antibody ELISA titer up to ~105

--High-throughput antibody screening platform: 10,000 clones per day

 

How to Order?

If you have any questions regarding our services or products, please feel free to contact us by E-mail: info@kmdbioscience.com or Tel: +86-400-621-6806;