Polyclonal Antibody Production in Rabbits

We use New Zealand white rabbits for the production of your polyclonal rabbit antibodies. It is the most used species to produce custom polyclonal antibodies and antisera. The advantages are a good amount of high-titer and high-affinity antibodies from the atisera at low costs. In addition rabbits are easy to handle and additional serum collections are easy to prepare and cost efficient. The final amount of antiserum is very good comparing with mice, rats or guinea pigs. Davids offers every step from the antigen synthesis/preparation to your final antibody. We help you with the decision, which kind of antigen is most suitable for your project and what kind of purification you need.

Antigen Preparation

Antigen Requirements

Immunization

Immunization Schedules

Antibody Purification

Available Purifications

Additional Services

Additional Services

Order

Order Process


Antigen Requirements

For the production of polyclonal rabbit antibodies a wide range of antigens can be used (Table 1).
When the purified (native) protein is available, it is usually the first choice for the antibody production. Purified proteins can be send at -20°C or -80°C to Davids wih cool packs or dry ice. We start the immunizations as soon as your antigen arrives. For proteins that are hard to purify, you can use our SDS-gel extraction service. In this case you load your protein to multiple bands of a SDS-gel and cut your specific gel bands from the gel. Send the gel pieces at room temperature to Davids. When the expression plasmid is available and time or equipment is not available for the expression and purification, you can use our protein expression service in E. coli.
For hard to express proteins, or proteins you only know the sequence from, we recommend to synthesize peptides that represent your protein. These peptides are conjugated to a carrier like KLH or BSA to improve the antigenicity prior to the immunization process.
The immunization of whole cells (bacteria, fungi, other eukaryotic cells) require a preparation to improve the presentation of the cells to the immune system. Please use Davids Cell Preparation KIT to improve the presentation of the surface from cells to the immune system. DNA as antigen is prepared by liposome introduction or by cationic detergents. Small molecules like drugs, toxins or haptens need to be prepared for immunization individually. Though if you have another antigen or when your antigen doesn't fit the requirements in the Table 1, you can contact us: info@davids-bio.com. For further information click on the desired antigen in the table.

Antigens for Rabbit Antibody Production
Antigen Amount
1 rabbit
Instructions Shipping
ProteinProtein0.5 mg
0.3 - 2 mg/ml
Customer provided proteins-20°C (ice packs) to -80°C (dry-ice) or lyophilized
Expression in E. coliProtein Expression-Expression Service by DavidsShip Expression Plasmid at -20°C
SDS-GelProtein (SDS-Gel)0.5 mg
3 - 8 gel bands
Protein Extraction from SDS-gel pieces. Cut the protein specific gel bands from the gel.Without addition of solutions at room temperature
PeptidePeptides1 - 2 mg
0.3 - 2 mg/ml
Customer provided peptides or Peptide synthesis and Conjugation Service by Davids.Lyophilized or with ice packs
CellCells106 - 107 cfu / immunizationPreparation with Davids Cell Immunization KIT for optimal presentation of the surface to the immune system.Please contact us when you want to send cells
DNADNA1 - 2 mg
0.3 - 2 mg/ml
DNA preparation by Davids.At room temperature or with ice packs
Conjugation of Small MoleculesSmall Molecules1 - 10 mg
0.3 - 2 mg/ml
Preparation of small molecules by DavidsPlease contact us when you want to send small molecules
For Affinity Purification we need additional >0.5 mg protein, 3 mg peptide or 108 cfu

Immunization Schedules

Comparison of titer development after immunization Each antigen profits for an optimized immunization schedule. With Davids established immunization schedules you profit from our experience. In addition to our well established immunization schedules we are able to follow your individual one.
Our 63-day Protocol includes 5 immunizations. After the third one (day 35) we take a test bleed and perform an ELISA titer determination. You are able to receive an aliquot of the test bleed and the results of the ELISA titer determination. After day 63 you receive the antiserum or we continue with a purification of the antiserum. The 63-day program is best for unknown antigens to achieve highest titers in a good time.

Davids 28-day SuperFast Immunization Protocol uses superior adjuvants and an optimized immunization schedule. The titers and antisera are comparable with our 63-day program (see picture). For peptides and small molecules Davids recommends the 63-day program.

Davids starts the immunizations as soon as the antigen arrives, usually at the same day. Table 2 shows an overview of our most popular polyclonal rabbit antibody services.

Immunization Protocols
SuperFast Well-Established LowDose Phospho-Specific
Cat.NoA055 (1 rabbit)A053 (1 rabbit)
A054 (2 rabbits)
A056 (1 rabbit)A060 (1 rabbit)
A061 (2 rabbits)
Fast RabbitTwo RabbitsSmall Amount of AntigenPhospho Antibody
Schedule28-day63-day63-day63-day
Schedule ProlongationNot PossiblePossibleNot PossiblePossible
Immunizations453 - 55
Test Bleed-day 35day 35day 35
Antiserum Amount40 - 90 ml40 - 90 ml40 - 90 ml-
Purification Options
PurificationPrecipitation (R001)
Affinity Purification (R007)
ProteinA (R008)
Precipitation (R001)
Affinity Purification (R007)
ProteinA (R008)
Precipitation (R001)
ProteinA (R008)
Affinity Purification included
Depletion included
Antigen Requirements
Suitable AntigensProtein
Protein (SDS-Gel)
Peptide
DNA
Cells
Small Molecules
Protein
Protein (SDS-Gel)
Peptide
DNA
Cells
Small Molecules
Protein
Protein (SDS-Gel)
Peptide
Cells
Small Molecules
Phospho Peptide
Antigen Needed0.5 mg Protein0.5 mg Protein < 0.5 mg Protein10 mg Peptide (we can synthesize it)
Antigen Concentration0.2 - 2 mg/ml0.2 - 2 mg/ml0.1 - 2 mg/ml1 - 10 mg/ml
Antigen BufferNon toxic bufferNon toxic bufferNon toxic bufferNon toxic buffer
PricePrice ListPrice ListPrice ListPrice List
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Our Immunization Schedules are well established and optimized for most customers. Though we are able to follow your immunization protocol or modify ours with additional intermediate bleeds, longer and additional immunizations and additional ELISA titer determinations.

Comparison of Immunization Schedules
Day 63 Day Antisera
1Preimmune serum, 0.5 ml
11. Immunization
142. Immunization
283. Immunization
35
  • Test serum, 1ml
  • ELISA-Titer determination
  • Optional: Test serum for the customer
424. Immunization
565. Immunization
63Antiserum, 40 - 90 ml
Get your offer to extend animal housing and the number of immunizations.
Day 28 Day SuperFast Antisera
1Preimmune serum, 1 ml
11. Immunization
72. Immunization
143. Immunization
214. Immunization
28Anti serum: 40 - 90 ml
Day Custom Protocol
XPreimmune serum, 1 ml
YImmunizations
ZIntermediate Bleed
AAntiserum: 40 - 90 ml

Rabbit Antiserum Purification

Davids superior purification services for your polyclonal rabbit antibodies lead to high quality and pure antibodies. Most common is the affinity purification, which we recommend for many applications like WesternBlot or IHC, to achieve specific antibodies that only recognize your antigen. This service is available for Proteins, Peptides and other antigens.
In addition we offer ProteinA purifications to receive a pure IgG fraction or depletions to get rid of unwanted antibodies that i. e. recognize tags, amino acid modifications or E. coli proteins.

Purification Options for Polyclonal Rabbit Antibodies
Affinity Purification ProteinA Cell Culture Purity Depletion
Cat.NoR007 (1 antiserum)
R007-2 (2 antisera)
R008 (1 antiserum)R003 (1 antiserum)R101 (anti-tag)
R102 (anti-E. coli)
Antigen Specific Affinity Purification SchemeProteinA Purification SchemeAffinity Purification SchemeAntibody Depletion Scheme
Antibody Purification Description
Duration5 days5 days5 - 10 days1 - 5 days
Product Information
ColumnAntigen bound to resinProteinA bound to resinAntigen bound to resintag/E. coli bound to resin
ProductSpecific AntibodiesWhole IgG FractionSpecific Antibodies
Low Endotoxin
Sterile
Fraction without unwanted Antibodies
Amount (approximately)0.5 - 10 mg5 - 30 mg0.1 - 10 mgDepends on Depleted Fraction
Applications
Optimal forWesternBlot
Immuno-Histo-Chemistry (IHC)
Fluorescent Activated Cell Sorting (FACS)
ELISA
Immuno-Histo-Chemistry (IHC)
in vivo Applications
Fluorescent Activated Cell Sorting (FACS)
Depends on Depleted Fraction
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Additional Services

To determine if a preimmune reaction exists before immunization Davids offers to determine in ELISA, which rabbit has the lowest reaction with your antigen. This can be important, when you want to minimize a cross-reactivity. Here we are able to immunize a rabbit that show only a small immune reaction to your antigen. Within the immunization schedule Davids can take additional small (0.5 ml) or large bleeds (15 ml) for ELISA determinations or for your applications.

Additional Services for Polyclonal Rabbit Antibodies
Cat.No. Description Amount Catalog Price
A057Additional Small Bleed0.5 - 1 mlorder form
A058Additional Large Bleed15 mlorder form
A059Additional Immunization1 Immunizationorder form
A070Preimmune Selection for the immunization of 1 Rabbit3 Preimmuneorder form
A705ELISA Raw Data incl. graphFrom 1 Bleedorder form
A706Animal Housing1 day, 1 rabbitorder form
N101Spleen Cell Isolation1 spleenorder form
N201PBMC Lymphocyte Isolation1 antiserumorder form
N301Bone Marrow Cell Isolation1 rabbitorder form
N401Lymph Node Cell Isolation1 rabbitorder form

Order Process

Davids keeps the order process very simple. You can send us your antigen to:

Davids Biotechnologie GmbH
Roentgenstrasse 3
93055 Regensburg
Germany

The immunizations usually start the same day the antigen arrives. To fasten up the immunization process and help us to connect your antigen with your order ensure you provide the following information:

  1. Your Antigen
  2. Concentration & Buffer Conditions of your Antigen
  3. Our Offer, our order form or your official PO
  4. VAT number (only EU Customers, not German Customers)

If you want to order peptide antibodies just send us your protein sequence and we suggest you peptides that represent your protein. For more information: Peptide Antibodies