About Us

Nebulabio is a globally recognized leader in custom antibody development, with over 20 years of specialized expertise in Polyclonal Antibody Production. Our service is dedicated to providing researchers, diagnostic developers, and biotech companies with a reliable, cost-effective solution for generating high-quality, high-titer polyclonal antisera against any antigen of interest. We excel in transforming diverse immunogens—from recombinant proteins and synthetic peptides to complex whole cells and small molecule haptens—into potent, multi-epitope recognition tools. Operating from our AAALAC-accredited facility, our team of experienced immunologists and protein scientists manages the entire production pipeline with an unwavering commitment to ethical standards, scientific rigor, and client success. We pride ourselves on delivering not just an antiserum, but a fully characterized, application-validated reagent that enhances the sensitivity and reliability of your critical assays.

☞Polyclonal Antibody Production Process

Nebulabio’s production process is a carefully optimized, multi-stage workflow designed to maximize immune response quality and ensure the highest yield of specific antibodies.

 Phase 1: Project Design & Immunogen Preparation 

• Strategic Antigen Consultation: We collaborate with you to design the optimal immunogen strategy. For peptides (<10 kDa) or haptens, we expertly conjugate them to carrier proteins (KLH, BSA, OVA) using site-specific chemistry (e.g., via cysteine or lysine residues) to maximize immunogenicity.

• Adjuvant Formulation: Your antigen is professionally emulsified with an appropriate adjuvant. We select from a range, including Freund's (Complete and Incomplete), aluminum-based salts (alum), or modern synthetic adjuvants (e.g., Titermax), based on the antigen type and host species to stimulate a robust and sustained Th2-biased humoral response.

 Phase 2: Ethical Immunization & Host Management 

• Host Species Selection: We recommend the optimal host—rabbit (standard), goat/sheep (for large volume), chicken (for IgY), or guinea pig—based on antigen homology, required serum volume, and project goals.

• Custom Immunization Protocol: Animals receive a primary subcutaneous or intramuscular injection, followed by a series of booster injections according to a tailored schedule, typically spanning 60-90 days. This extended period allows for thorough B-cell clonal expansion and affinity maturation.

• Serum Titer Monitoring: Pre-immune serum is collected, and regular test bleeds are taken during the immunization course. Serum is analyzed via ELISA to quantitatively monitor the rise of antigen-specific antibody titers, ensuring booster injections are timed for maximum effect.

 Phase 3: Terminal Bleed, Serum Collection & Processing 

• Upon confirmation of a high, plateaued titer, a final, large-volume terminal bleed is performed humanely by trained personnel.

• The blood is allowed to clot, and the serum is separated via centrifugation. The crude antiserum is then clarified through filtration, aliquoted, and stored at -20°C or below.

Phase 4: Purification & Fractionation Options 

We offer tiered purification services to match your application's needs:

• Crude Antiserum: The unpurified serum, containing all serum proteins including the specific IgG.

• Total IgG Purification: Using affinity chromatography with Protein A (for rabbit, mouse, human) or Protein G (for broader species including goat, sheep, cow), we isolate the total IgG fraction. This removes the bulk of serum albumin and other contaminants, significantly reducing background in assays.

• Antigen-Specific Affinity Purification: The premium option. The antiserum is passed over a column with your immobilized antigen. This isolates only the antibodies specific to your target, yielding a product of exceptional purity and specificity, ideal for applications like blocking studies or labeling where cross-reactivity must be minimized.

 Phase 5: Comprehensive Quality Control 

Every batch undergoes rigorous QC before release:

• Titer & End-Point Dilution Analysis by ELISA.

• Specificity Verification via Western Blot against relevant cell lysates or tissues.

• Application Testing (e.g., IHC on known positive/negative tissue sections) can be performed upon request.

• Concentration Measurement (A280).

• Purity Assessment by SDS-PAGE.

Figure1. Approaches used to produce current clinically approved Polyclonal Antibody.

☞Why Choose Nebulabio’s Polyclonal Antibody Service?

Selecting our service provides distinct advantages in quality, reliability, and scientific partnership.

• Unmatched Experience & High Success Rates: Our two decades of specialization mean we have successfully produced antibodies against thousands of antigens, including notoriously difficult targets like small peptides, conserved proteins, and post-translationally modified antigens. Our optimized protocols maximize your chance of success.

• Full Customization & Expert Guidance: We offer true customization, not pre-set packages. Our scientists provide expert consultation on every critical choice—from host species and adjuvant selection to peptide design and conjugation chemistry—tailoring the project to your specific antigen and intended use.

• Ethical Excellence & Animal Welfare: Our AAALAC accreditation underscores our commitment to the highest standards of animal care and use. We adhere to strict ethical guidelines, ensuring humane treatment throughout the immunization process, which also contributes to a healthier immune response and higher-quality antiserum.

• Rigorous Quality Control & Validation: We don't just deliver serum; we deliver characterized data. Our standard QC includes titer and specificity validation, and we offer application-specific testing. This ensures the antibody performs as expected in your hands, saving you valuable time and resources.

• Scalable Production & Purification Expertise: Whether you need 10 mL of crude serum or grams of affinity-purified IgG, we have the capacity and expertise. Our in-house purification team handles everything from simple IgG isolation to complex antigen-affinity columns.

• Dedicated Project Management: You are assigned a single point of contact—a project manager who provides regular updates, detailed reports (including titer curves), and is readily available to answer questions, ensuring a transparent and collaborative experience from start to finish.

☞Applications of Polyclonal Antibodies

The unique advantages of polyclonal antisera—high sensitivity, multi-epitope recognition, and robustness—make them essential for numerous techniques:

• Western Blotting: The most common application. Their ability to bind multiple epitopes often results in stronger signals and higher sensitivity for detecting proteins in complex lysates, even if the protein is partially denatured.

• Immunohistochemistry (IHC) & Immunofluorescence (IF): Widely used for visualizing protein localization in fixed tissues and cells. Polyclonals can provide intense, clear staining and are often more forgiving of tissue fixation variations than some monoclonals.

• Enzyme-Linked Immunosorbent Assay (ELISA): Frequently employed as the capture antibody in sandwich ELISA formats due to their high affinity and ability to bind the target antigen at multiple sites, ensuring efficient capture from solution.

• Immunoprecipitation (IP) & Co-Immunoprecipitation (Co-IP): The method of choice for "pulling down" native proteins and their interaction complexes. The formation of a large antigen-antibody lattice facilitates efficient precipitation.

• Chromatin Immunoprecipitation (ChIP): Used to precipitate protein-DNA complexes, essential for studying gene regulation, transcription factor binding, and epigenetic marks.

• Blocking & Neutralization Studies: In functional assays, polyclonal antisera are effective at blocking receptor-ligand interactions, neutralizing cytokines, pathogens, or toxins due to their ability to target multiple sites on the target molecule.

☞Nebulabio’s Service Workflow

Our client-centric workflow is designed for clarity, efficiency, and partnership.

 Nebulabio Service Process Flowchart: 

Partner with Nebulabio’s experts for your polyclonal antibody needs. Our comprehensive service combines deep technical expertise with a commitment to quality, delivering a powerful, reliable detection tool that strengthens your research or diagnostic pipeline.