Anti-Arabinogalactan-protein [LM30] monoclonal antibody
Invented by Prof Paul Knox from PlantProbes
Invented at University of Leeds
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Info
| Catalogue Number | 157934 |
| Antigen/Gene or Protein Targets | Arabinogalactan-protein |
| Relevance |
The Gsp-1 genes of wheat are proposed to encode precursor proteins that are post-translationally processed to give two components: the arabinogalactan peptide (comprising 15 amino acids) and the grain softness protein (GSP-1) (the major form comprising 113 amino acids with 10 cysteine residues) (Elmorjani et al., 2013). However, it has not been directly demonstrated that the AGP is derived from Gsp-1 genes. Gsp-1 genes appear to be ubiquitous in grasses, being identified in 65 species from the five major grass subfamilies, with over 20 different Gsp-1 alleles being reported (Wilkinson et al., 2013). The GSP protein belongs to a large family of seeds storage proteins called the “prolamin superfamily” (Shewry and Halford, 2002) and has been predicted to have a similar structure to other members of this family, including the 2S storage albumins which occur in dicotyledonous seeds and the puroindolines (Pins) of cereal seeds (Elmorjani et al., 2013). |
| Host | Rat |
| Immunogen | Wheat AGP in complete Freund's adjuvant. |
| Positive Control | IgM |
| Myeloma Used | IR983F |
| Notes |
Isolated subsequent to immunization with a preparation of arabinogalactan-protein (AGP) obtained from wheat grain. LM30 recognizes a glycan epitope of the wheat AGP and the recognition is highly sensitive to the action of arabinofuranosidase. AGP was isolated from Cadenza white flour as described in Tryfona et al. (2010), freeze-dried and stored at −80 °C. Two male Wistar rats were injected with ∼100 μg each of wheat AGP in complete Freund's adjuvant administered sub-cutaneously on day 0, with the same amount administered with incomplete Freund's adjuvant on days 35, 70 and 98. On day 147, a rat was given a pre-fusion boost of 100 μg immunogen in 1 ml PBS by intraperitoneal injection. Spleen lymphocytes were isolated three days later and fused with rat myeloma cell line IR983F (Bazin, 1982). Hybridoma cell lines were selected by ELISA using the wheat AGP as antigen. |
| Research Area | Plant Science |
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