Phosphorylated Monoclonal & Polyclonal Antibody Services

KMD Bioscience has extensive experience producing customized phosphorylated monoclonal and polyclonal antibodies. We can provide specific phosphorylation sites antibody production service and the success rate is up to 100% till now. 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 very important post-translational modification (PTM) in cells, mainly involved in the regulation of 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 and antibody purification and pairing services.

 

Phosphorylated Antibody Development Service Protocol: 

Procedure

Description

Deliverable

Antigen Design & Synthesis

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

2. KMD Bioscience design and synthesis phosphorylated and non-phosphorylated antigens (optional)

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

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

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

Monoclonal Abs Development

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

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

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

*2-5 hybridoma cell lines

*2-5 mg purified antibody

*ELISA report

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

*Western Blot validation report (optional)

Polyclonal Abs Development

1. 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

2. Serum collection and ELISA titer detection

3. Antigen affinity purification (recommended) or protein A purification

4. Antibodies are verified using dilution factor (customer samples are required)

*1-2ml pre-immune serum

*1-5 mg purified antibody

*ELISA report

*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