About Us

Welcome to Nebulabio’s specialized Custom Rabbit Polyclonal Antibody Production Service, a cornerstone of our comprehensive antibody development portfolio. Rabbits are widely regarded as the premier host for generating high-quality polyclonal antisera due to their robust immune response, relatively large serum yield, and ability to produce antibodies against a diverse array of antigens, including those that are highly conserved or of low immunogenicity in other species. Our service is dedicated to transforming your specific antigen—whether it’s a recombinant protein, synthetic peptide, or small molecule hapten—into a potent, well-characterized polyclonal antiserum. Managed by a team of expert immunologists within our AAALAC-accredited facility, we ensure an ethical, scientifically rigorous, and fully customized production process. We deliver more than just serum; we provide a reliable, high-performance detection tool validated for your specific research or diagnostic application.

☞Rabbit Polyclonal Antibody Production Process

Nebulabio’s production process is a meticulously optimized, stage-gated workflow designed to elicit a strong, specific immune response and yield antiserum of the highest possible quality and consistency.

 Phase 1: Antigen Design & Immunogen Preparation 

• Expert Strategy Session: We work closely with you to design the optimal immunogen. For peptides (typically 10-15 amino acids) or haptens, our chemists perform professional conjugation to keyhole limpet hemocyanin (KLH) or bovine serum albumin (BSA) carriers using controlled chemistry (e.g., via cysteine residues or lysine amines) to maximize immunogenicity.

• Adjuvant Formulation: The antigen is emulsified with an appropriate adjuvant. We typically use Freund's Adjuvant (Complete for primary immunization, Incomplete for boosts) for robust, long-lasting responses, or offer alternatives like Titermax Gold for specific project needs.

 Phase 2: Immunization & Immune Monitoring 

• Ethical Host Management: New Zealand White rabbits are housed under strict, ethical guidelines in our accredited facility. Each project uses at least two rabbits to account for individual immune response variability.

• Custom Immunization Schedule: Rabbits receive a primary subcutaneous multi-site injection, followed by 4-5 booster injections over an 8-12 week period. This extended schedule promotes thorough B-cell clonal selection and affinity maturation.

• Titer Analysis: Pre-immune serum is collected. After the 3rd boost and each subsequent boost, test bleeds are taken. Serum is analyzed by indirect ELISA to quantitatively track the rise of antigen-specific antibody titers, ensuring boosts are administered at the peak of immune response.

 Phase 3: Terminal Bleed & Serum Processing 

• Once antibody titers have reached a high plateau (typically >1:64,000 by ELISA), a final, large-volume terminal bleed is performed humanely via cardiac puncture under anesthesia.

• The blood is allowed to clot, and the serum is separated by centrifugation. The crude antiserum is then filtered (0.45 µm), aliquoted to prevent freeze-thaw cycles, and stored at -20°C or -80°C.

 Phase 4: Purification & Processing Options 

We offer multiple purification tiers to suit your application and budget:

• Crude Antiserum: The unpurified, total serum containing your specific IgG alongside other serum proteins.

• Total IgG Purification (Standard): Using Protein A affinity chromatography, which specifically binds rabbit IgG, we isolate the total IgG fraction. This removes ~90% of contaminating proteins (like albumin), significantly reducing background noise in assays. This is our most popular and cost-effective option.

• Antigen-Specific Affinity Purification (Premium): The antiserum is passed over a column containing your immobilized antigen. This isolates only the antibodies specific to your target, yielding a product of exceptional purity and specificity, ideal for highly sensitive applications or conjugation.

 Phase 5: Comprehensive Quality Control (QC) 

Every batch of antiserum undergoes rigorous QC:

• Titer Verification: Final ELISA titer confirmation.

• Specificity Analysis: Western blot against a relevant cell or tissue lysate to confirm recognition of the target at the expected molecular weight and to assess potential cross-reactivity.

• Application-Specific Testing (Optional): Performance testing in IHC, IP, or other client-specified assays.

• Concentration & Purity: Determination of IgG concentration (A280) and assessment by SDS-PAGE.

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

☞Nebulabio’s Service Workflow

 Step 1: Project Consultation & Proposal 

Contact us with details about your antigen and intended application. We schedule a technical discussion to review your needs and provide a detailed, itemized proposal with timeline and cost.

 Step 2: Project Initiation & Antigen Submission 

Upon agreement, we send a project initiation package. You then ship your purified antigen (>1 mg for protein, >5 mg for peptide recommended) to our secure facility.

 Step 3: Immunization Program with Regular Updates 

Our team formulates the immunogen and begins the rabbit immunization protocol. You receive scheduled progress reports, including serum titer data presented in clear graphical format, allowing you to monitor the immune response in real time.

 Step 4: Final Collection, Purification & QC 

At the completion of the immunization schedule, we perform the terminal bleed, process the serum, and execute the chosen purification method. The final product then enters our QC pipeline.

 Step 5: Final Delivery & Support

We ship the final, QC-approved product: the purified rabbit polyclonal antibody, formulated in a standard buffer (e.g., PBS) or as lyophilized powder. The shipment includes:

• A comprehensive Certificate of Analysis with all QC data.

• A small aliquot of pre-immune serum (when available) as a critical negative control.

• Detailed technical information on storage and handling.

 Nebulabio Service Process Flowchart: 

☞Why Choose Our Service?

Partnering with us for your custom rabbit polyclonal antibodies offers distinct advantages in quality, reliability, and scientific partnership.

• Specialized Rabbit Expertise: Our protocols are optimized specifically for the rabbit immune system. We understand the nuances of adjuvant selection, boost timing, and peptide design to elicit the strongest possible response against your target, leading to higher average titers and success rates.

• Fully Customized, Not Pre-Packaged: We treat every project as unique. Our scientists provide expert guidance on peptide sequence selection, conjugation strategy, and immunization schedule tailored to your specific antigen’s properties and your final application requirements.

• Uncompromising Quality Control & Validation: We believe an antibody is only as good as its validation data. Our standard QC goes beyond simple titering to include specificity verification by Western blot. This commitment ensures the antiserum you receive will perform reliably, saving you valuable time and resources in assay optimization.

• Ethical Production & Animal Welfare Commitment: Our AAALAC accreditation is a testament to our highest standards of animal care and welfare. Ethical treatment is not only a moral imperative but also contributes to less stressed, healthier animals, which is known to promote a more robust and consistent immune response.

• Scalable Production & Purification Expertise: From small research projects needing a few milliliters to large-scale diagnostic development requiring grams of purified IgG, we have the capacity and expertise. Our in-house protein purification team handles everything from standard Protein A columns to complex antigen-specific affinity purification.

• Dedicated Project Management & Transparency: You are assigned a dedicated project manager—a single point of contact who provides regular updates, detailed reports (including raw ELISA data if requested), and is readily available to answer questions, ensuring a collaborative and transparent experience.

☞Applications of Rabbit Polyclonal Antibodies

Rabbit polyclonal antisera are versatile, high-sensitivity reagents critical for a wide range of biomedical research and diagnostic techniques.

• Western Blotting: The most common application. The multi-epitope recognition of polyclonals often provides stronger signals and higher sensitivity for detecting proteins in complex lysates, and they are more tolerant of protein denaturation compared to some monoclonal antibodies.

• Immunohistochemistry (IHC) & Immunofluorescence (IF): Rabbit polyclonals are excellent for visualizing protein localization in fixed tissues and cells. Their high affinity and signal amplification can reveal low-abundance targets, and they are less likely to cross-react with endogenous mouse IgG in mouse tissue studies, reducing background.

• Immunoprecipitation (IP) & Co-Immunoprecipitation (Co-IP): Often the preferred choice for "pulling down" native proteins and their interacting partners. The ability to form a large antigen-antibody lattice with multiple epitopes facilitates efficient precipitation of the target complex.

• Enzyme-Linked Immunosorbent Assay (ELISA): Frequently used as the capture antibody in sandwich ELISA formats. Their high affinity and ability to bind multiple sites on the target antigen ensure efficient capture from solution.

• Blocking & Neutralization Studies: In functional assays, polyclonal antisera can effectively block receptor-ligand interactions, neutralize cytokines, or inhibit enzyme activity due to their ability to bind and sterically hinder multiple regions on the target molecule.

Elevate your research with a trusted detection partner. Contact Nebulabio today to initiate your Custom Rabbit Polyclonal Antibody project and leverage our expertise to obtain a high-quality, validated reagent tailored to your precise needs.