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Demonstration of Immunoprecipitation & Flow Cytometry Specific Antibody Production

Top panel illustrates the strategy to promote in vitro folding and stabilizing of hydrophilic domains in a target protein or epitope sequence. Bottom panel, left, shows the immunoprecipitation by anti-claudin-1 immunoprecipitation specific antibody from Pichia yeast cell lysate expressing claudin-1 protein. Right, shows the magnetic bead sorting with anti-claudin-18-ECL flow cytometry specific antibody of HEK293 cells expressing claudin-18 protein.

Immunoprecipitation & Flow Cytometry Specific Antibody Production

$2,150.00 – $8,950.00

Antibody: Polyclonal and Monoclonal Antibody

Concentration: 0.25 mg/ml of purified IgG

Application: Immunoprecipitation & Flow Cytometry

Reactivity: Human, Mouse, Rat

BiCell Scientific Inc has developed Immunoprecipitation & Flow Cytometry-specific recognition antibodies with novel in vitro folding and stabilizing technique to lock the conformational state of hydrophilic domains in an epitope or target protein in order to pull down proteins in aqueous environment from immunoprecipitated samples or live cells.

Immunoprecipitation & Flow Cytometry requires antibodies that can detect protein foldings in aqueous environment. Conventional antibodies lack the binding affinity specific to the hydrophilic domains that are stabilized by interaction with water molecules, which limits the antibody’s ability to pull down target proteins or live cells in immunoprecipitation or cell sorting applications.

BiCell Scientific Inc has developed an in vitro folding and stabilizing technique to lock the conformational state of hydrophilic domains in a target protein in order to pull down proteins in aqueous environment from immunoprecipitated samples or live cells. BiCell Scientific Inc is able to raise polyclonal and monoclonal antibodies against a target protein and its hydrophilic domains. Such antibodies can facilitate protein purification by immunoprecipitation or cell sorting by flow cytometry.

Polyclonal Antibody Workflow Chart

Steps Timeline (weeks) Deliverables/Internal testing milestone QC standard
Phase I: Antigen preparation

(peptide antigen,13-19 aa length, will be designed against target protein, synthesized, in vitro folded, stabilized and conjugated to KLH)

 2 weeks / COA

 

 

 

 
Phase II: Animal immunization

(3 Sprague Dawley rats or 5 Swiss Webster mice or 2 New Zealand rabbits will be immunized with antigen; conventional protocol – three injections)

 

 

 8-10 weeks 100 ul antiserum ELISA after 3rd injection
Phase III: Antiserum collection

(antiserum will be collected from one best responder animal according to ELISA report)

 

 

 1-2 weeks 1 ml to 50 ml antiserum ELISA from production bleed
Phase IV: Affinity Chromatography and Depletion Chromatography

(antiserum will be purified with affinity column conjugated with the antigen peptide)

 

 1-2 weeks 1 mg of purified polyclonal IgG IP/Flow cytometry

Monoclonal Antibody Workflow Chart

Steps Timeline (weeks) Deliverables/Internal testing milestone QC standard
Phase I: Antigen preparation

(peptide antigen,13-19 aa length, will be designed against target protein, synthesized, in vitro folded, stabilized and conjugated to KLH)

 2 weeks / COA

 

 

 

 
Phase II: Animal immunization

(3 Sprague Dawley rats or 5 Swiss Webster mice or 2 New Zealand rabbits will be immunized with antigen; conventional protocol – three injections)

 

 

 8-10 weeks 100 ul anti-serum ELISA after 3rd injection
Phase III: Cell fusion and screening

(fusion of mouse myeloma cells with splenocytes from one best responder animal to generate hybridoma and perform antigen based hybridoma selection, enriching, and plating)

 

 

 4-6 weeks Supernatant from 96 wells ELISA from supernatant
Phase IV: Expansion and antibody purification

(customer choosing one clone; expanding clone and purifying antibodies with affinity chromatography)

 

 6-8 weeks One hybridoma cell line (to customer); purified monoclonal antibody from the selected hybridoma cell line (1mg)

(BiCell Scientific will bank all remaining clones; customer can request additional clones by paying a service fee for hybridoma recovery)

 IP/Flow cytometry

 

For Research Use Only. Not for use in clinical diagnostics.

Additional information

Antibody Deliverable

1 mg of Immunoprecipitation & Flow Cytometry Specific Polyclonal Antibody Production, 1 mg of Immunoprecipitation & Flow Cytometry Specific Monoclonal Antibody Production
Host Species

Rat, Mouse, Rabbit

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"I am really impressed with your approach. We tried multiple times previously to create monoclonal and polyclonal antibodies to claudin-2 and MLCK1. We have had limited success generating polyclonals and no success generating monoclonals. You have generated outstanding monoclonals to both. I look forward to continuing to work with you."

Jerrold R. Turner, M.D., Ph.D.

Brigham and Women’s Hospital | Harvard Medical School

"The polyclonal antibody you generated for KIAA0408 is stunning! KIAA0408 is a novel cilium molecule that has never been studied. So, clearly there will be a lot of demand for it as we have discovered a very interesting finding and the story will be published in a high impact journal. I am strongly inclined to generate monoclonal antibody for this protein too and we should think about patenting it."

Univ.-Prof. Jay Gopalakrishnan PhD

Heinrich-Heine-Universität | Universitätsklinikum Düsseldorf

"Your ARL13B antibody works beautifully!!! We’re so happy to have a cilia-specific antibody made in rat! I can send you high resolution images to be posted on your website."

Julie Craft Van De Weghe, PhD

School of Medicine | University of Washington

"The assay is a homophilic interaction mediated cell adhesion on purified protein (in this case, immobilized purified Pcdhga9 to Pcdhga9 expressed on cell surface). Compared to control, cell adhesion is reduced in the presence of Pcdhga9 monoclonal antibody supernatants!"

Divyesh Joshi, PhD

School of Medicine | Yale University

"The rabbit hybridoma supernatants of anti-APOBEC3 project are tested positive by ELISA, and we are very happy about it! We previously tried a company, Abclone. Their Project "A" has immune response that is <10,000 titer in antiserum, which would explain why there is no positive mAb after fusion. Their project "B" didn't have any immune response in rabbit."

Harshita B Gupta, PhD.

School of Medicine | UT Health San Antonio

"We have tested anti mouse T cell antiserum samples from both rabbits you sent to us.

They worked very well! Thank you!"

Victoria Gorbacheva, PhD.

School of Medicine | Cleveland Clinic

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