Rabbit Antibody Production

Antisera & Affinity Purified Antibodies

For the manufacturing of your polyclonal rabbit antibodies, we select New Zealand white rabbits. It is the most often chosen species for the production of custom polyclonal antibodies and antisera. The benefits include a large quantity of high-titer and high-affinity antibodies from antisera at a reasonable cost. Furthermore, rabbits are easy to handle, and subsequent serum samples are simple to prepare and cost effective. When compared to mice, rats, or guinea pigs, the ultimate amount of antiserum is very good. Davids provides every stage of the process, from antigen synthesis/preparation to your finalized antibody. We assist you in determining which antigen is most suited to your project and what type of purification is required.

Antigen Requirements

A variety of antigens can be used to generate polyclonal rabbit antibodies (Table 1). When accessible, pure (native) protein is generally the first choice for antibody production. Purified proteins can be sent to Davids at -20°C or -80°C using cold packs or dry ice. As soon as your antigen arrives, we begin with the vaccinations. You can utilize our SDS-gel extraction service for difficult-to-purify proteins. In this case, you load your protein onto various bands of an SDS-gel and then cut the specific gel bands from the gel. Send the gel pieces to Davids at room temperature.

When an expression plasmid is available but there is insufficient time or equipment for expression and purification, you can use our protein expression service in E. coli or use peptides that represent your protein. For proteins, that are difficult to express or proteins you only know the sequence from, we recommend to synthesize peptides that represent your protein. Prior to vaccination, these peptides are conjugated to a carrier such as KLH or BSA to increase antigenicity.

Immunization of complete cells (bacteria, fungus, and other eukaryotic cells) need a preparation to enhance the cells' presentation to the immune system. Please utilize Davids Cell Preparation KIT to increase cell surface presentation to the immune system. Liposomes or cationic detergents are used to prepare DNA as an antigen. Small compounds, like as drugs, toxins, or haptens, must be specially prepared for immunization. However, if you have another antigen or if your antigen does not meet the standards in Table 1, please contact us with our contact form: Antibody Creator. Click on the selected antigen in the table for further details.

	Antigen	Amount	Instructions	Shipping
	Protein	0.5 - 1 mg 0.3 - 2 mg/ml	Proteins supplied by customer	-20°C (ice packs) to -80°C (dry-ice) or lyophilized
	Protein Expression	-	Davids expression service	Provide us your protein sequence
	Protein (SDS-Gel)	0.5 - 1 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
	Peptides	1 - 2 mg 0.3 - 2 mg/ml	Customer provided peptides or Peptide synthesis and Conjugation Service by Davids.	Lyophilized or with ice packs
	Cells	106 - 107 cfu / immunization	Preparation with Davids Cell Immunization KIT for optimal presentation of the surface to the immune system.	Please contact us when you want to send cells
	DNA	1 - 2 mg 0.3 - 2 mg/ml	DNA preparation by Davids.	At room temperature or with ice packs
	Small Molecules	1 - 10 mg 0.3 - 2 mg/ml	Preparation of small molecules by Davids	Please contact us when you want to send small molecules
For Affinity Purification we need additional 0.5 - 2 mg protein, 3 mg peptide or 108 cfu

Immunization Schedules

Each antigen profits from a well-planned vaccination program. You benefit from our experience when you use Davids established immunization schedules. We can follow your personalized immunization program in addition to our well-established ones. Five immunizations are included in our 63-day Protocol. After the third (day 35), we take a test bleed and determine the ELISA titer. You can obtain an aliquot of the test bleed along with the ELISA titer determination results. After day 63, you will either receive the antiserum or we will proceed with the antiserum purification. The 63-day strategy is ideal for unknown antigens in order to produce the greatest titers in the shortest amount of time.
Davids begins with the immunizations as soon as the antigen arrives in our lab, which is usually the same day. Table 2 lists our most popular polyclonal rabbit antibody services.

	SuperFast	Well-Established	LowDose	Phospho-Specific
Cat.No	A055 (1 rabbit)	A053 (1 rabbit) A054 (2 rabbits)	A056 (1 rabbit)	A060 (1 rabbit) A061 (2 rabbits)
Schedule	28-day	63-day	63-day	63-day
Schedule Prolongation	Not Possible	Possible	Not Possible	Possible
Immunizations	4	5	3 - 5	5
Test Bleed	-	day 35	day 35	day 35
Antiserum Amount	40 - 90 ml	40 - 90 ml	40 - 90 ml	-
Purification Options
Purification	Precipitation (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 Antigens	Protein 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 Needed	0.5 mg Protein	0.5 mg Protein	< 0.5 mg Protein	10 mg Peptide (we can synthesize it)
Antigen Concentration	0.2 - 2 mg/ml	0.2 - 2 mg/ml	0.1 - 2 mg/ml	1 - 10 mg/ml
Antigen Buffer	Non toxic buffer	Non toxic buffer	Non toxic buffer	Non toxic buffer
Price	Price List	Price List	Price List	Price List
Get your free quotation now

Our Immunization Schedules have been thoroughly established and optimized for the majority of our clients. We can follow your vaccination protocol or adapt ours with more intermediate bleeds, extended and additional immunizations, and ELISA titer determinations.

Day	63 Day Antisera
1	Preimmune serum, 1 ml
1	1. Immunization
14	2. Immunization
28	3. Immunization
35	Test serum, 0.5 ml ELISA-Titer determination Optional: Test serum for the customer
42	4. Immunization
56	5. Immunization
63	Antiserum, 40 - 90 ml
∞	Get your offer to extend animal housing and the number of immunizations.

Day	28 Day SuperFast Antisera
1	Preimmune serum, 1 ml
1	1. Immunization
7	2. Immunization
14	3. Immunization
21	4. Immunization
28	Anti serum: 40 - 90 ml

Day	Custom Protocol
X	Preimmune serum, 1 ml
Y	Immunizations
Z	Intermediate Bleed
A	Antiserum: 40 - 90 ml

Rabbit Antiserum Purification

The exceptional purification services provided by Davids for your polyclonal rabbit antibodies result in high quality and pure antibodies. The most popular option is affinity purification, which we suggest for many applications such as WesternBlot or IHC to obtain specific antibodies that identify just your antigen. This service is available for proteins, peptides, and other antigens. In addition, we provide ProteinA purifications to get a pure IgG fraction, as well as depletions to remove undesired antibodies that detect tags, amino acid changes, or E. coli proteins.

	Affinity Purification	ProteinA	Cell Culture Purity	Depletion
Cat.No	R007 (1 antiserum) R007-2 (2 antisera)	R008 (1 antiserum)	R003 (1 antiserum)	R101 (anti-tag) R102 (anti-E. coli)
	Antigen Specific Antibodies:	Whole IgG Fraction:	Antigen Specific Antibodies:	Anti-tag Antibody Removal:
	Tag: Antigen Specific Antibody: Unspecific Antibody: Tag Antibody: Antigen:
Duration	5 days	5 days	5 - 10 days	1 - 5 days
Product Information
Column	Antigen bound to resin	ProteinA bound to resin	Antigen bound to resin	tag/E. coli bound to resin
Product	Specific Antibodies	Whole IgG Fraction	Specific Antibodies Low Endotoxin Sterile	Fraction without unwanted Antibodies
Amount (approximately)	0.5 - 10 mg	100 - 200 mg (from 30 ml Antiserum)	0.1 - 10 mg	Depends on Depleted Fraction
Applications
Optimal for	WesternBlot 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
Price	Price List	Price List	Price List	Price List
Get your free quotation now

Additional Services

To verify if a preimmune reaction occurs prior to vaccination, Davids recommends utilizing ELISA to discover which rabbit has the lowest reactivity with your antigen. This is helpful when trying to lower cross-reactivity. Here, we may immunize a rabbit that has just a minor immunological response to your antigen. Davids can collect extra small (0.5 ml) or big (15 ml) bleeds throughout the vaccination plan for ELISA determinations or for your applications.

Cat.No.	Description	Amount	Catalog Price
A057	Additional Small Bleed	0.5 - 1 ml	order form
A058	Additional Large Bleed	15 ml	order form
A059	Additional Immunization	1 Immunization	order form
A070	Preimmune Selection for the immunization of 1 Rabbit	3 Preimmune	order form
A705	ELISA Raw Data incl. graph	From 1 Bleed	order form
A706	Animal Housing	1 day, 1 rabbit	order form
N101	Spleen Cell Isolation	1 spleen	order form
N201	PBMC Lymphocyte Isolation	1 antiserum	order form
N301	Bone Marrow Cell Isolation	1 rabbit	order form
N401	Lymph Node Cell Isolation	1 rabbit	order form

Order Process

Davids keeps the ordering procedure as easy as possible. Please send your antigen to:

Davids Biotechnologie GmbH
Roentgenstrasse 3
93055 Regensburg
Germany

Immunizations usually begin on the same day as the antigen arrives. Please submit the following information to assist us correlate your antigen with your order and accelerate the process:

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 required for German customers)

Get your quote now with our Antibody Creator. When you prefer to order peptide antibodies, send us your protein sequence and we will recommend peptides that represent your protein. For further information, please visit: Peptide Antibodies