Rabbit is an excellent choice of host. Traditionally used, cost effective and with an abundance of secondary antibodies rabbit-antibodies are suitable for most projects. What is more, for anti-peptide antibody projects, where we have designed and produced the peptide, we will guarantee that the resulting antiserum will contain antibodies that target the peptide.
Of course, you can fully customise your project, but for your convenience we have outlined a typical project.
An immunogen is required for the antibody production. This can be a protein or other molecule that you provide, or it can be produced here. In the latter case, either the entire protein can be obtained using a recombinant technique, or a peptide is designed and synthesized. Using peptides as antigens is the most frequently used strategy. It should be noted that the more effort that is put into this step, the better the odds for the successful outcome of your project. Other labs may choose to ignore this fact, but the production of high quality peptides for antibody production may very well be pivotal for the success of your project.
If you will be supplying the immunogen, there are a few things to keep in mind;
One or two rabbits are immunised using our standard protocol. For each rabbit pre-immune serum is collected prior to immunisations, and three bleeds are made over the course of the immunisation schedule. Each 20 ml bleed contains in the order of 100 mg total IgG.
The immunisation protocol is an open-ended schedule, meaning that termination requires your explicit request. Managing your project will ensure that you obtain antibody in the required amounts, while keeping costs down. This means collecting a sufficient volume of antiserum and then terminating the project as soon as possible.
The crude rabbit antiserum often works very well without any purification. If you wish, we can purify serum and collect either the total IgG, or the fraction of antigen specific antibodies. Typically 20 ml serum is used for such a purification, and the typical yield is 2-5 mg of antigen specific antibody.
Examples of references of customers using our custom rabbit antibody services include;
H. Han, D. Xu, C. Liu, H-E. Claesson, M. Björkholm, J. Sjöberg
Interleukin-4-Mediated 15-Lipoxygenase-1 Trans-Activation Requires UTX Recruitment and H3K27me3 Demethylation at the Promoter in A549 Cells
PLOS, January 20, 2014 doi: 10.1371/journal.pone.0085085
R. Bahtz, J. Seidler, M. Arnold, U. Haselmann-Weiss, C. Antony, W. D. Lehmann and I. Hoffmann
GCP6 is a substrate of Plk4 and required for centriole duplication
Journal of Cell Science, February 2, 2012 doi: 10.1242/jcs.093930
H. H. Qi, P. P. Ongusaha, J. Myllyharju, D. Cheng, O. Pakkanen, Y. Shi, S. W. Lee, J. Peng & Y. Shi.
Prolyl 4-hydroxylation regulates Argonaute 2 stability.
Nature, 18 September, 2008, 455, 421-424
M. W. Hornef, T. Frisan, A. Vandewalle, S. Normark, and A. Richter-Dahlfors.
Toll-like Receptor 4 Resides in the Golgi Apparatus and Colocalizes with Internalized Lipopolysaccharide in Intestinal Epithelial Cells.
Journal of Experimental Medicine, February 25, 2002, vol. 195 no. 5 559-570.
L. Camper, U. Hellman, E. Lundgren-Åkerlund
Isolation, Cloning, and Sequence Analysis of the Integrin Subunit α10, a β1-associated Collagen Binding Integrin Expressed on Chondrocytes.
The Journal of Biological Chemistry, August 7, 1998, vol 273, 20383-20389.
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