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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 hydrophobic domains in an epitope or target protein similar to naturally folded proteins in immunoprecipitated samples or live cell membrane.

Immunoprecipitation & Flow Cytometry detect naturally folded proteins under physiologic conditions. Conventional antibodies lack the binding affinity specific to the natural conformation of a target protein that is stabilized by internal hydrophobic domains, which limits the antibody’s ability to pull down target proteins or cells in physiologic buffers.

BiCell Scientific Inc has developed an in vitro folding and stabilizing technique to lock the conformational state of hydrophobic domains in an epitope or target protein similar to naturally folded proteins in immunoprecipitated samples or live cell membrane. BiCell Scientific Inc is able to raise polyclonal and monoclonal antibodies against a naturally folded target protein or an epitope sequence. 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

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