Antibodies that recognise a Tc1 Hsa21 specific protein RRP1 One of the anti RRP1 antibodies, which was purified towards peptide B, Inhibitors,Modulators,Libraries recognised a 50 kDa band on western blots of Tc1 total brain proteins, steady with all the predicted molecular weight of RRP1. A very similar band was not observed in non transchro mosomic management mice, indicating that this antibody might exclusively react with human RRP1. RRP1 peptide sequence B is one of a kind towards the human protein and is not observed in mouse RRP1. In addition for the Tc1 distinct band a number of weaker extra bands had been observed in samples of Tc1 and non Tc1 total brain proteins. They are likely to represent non precise inter action from the polyclonal antibody with other brain professional teins.
Despite the relative specificity of your 9644 B antibody on western blot, a comparable pattern and intensity of staining was observed on Tc1 and non transchromo somic management mouse entire brain sections, intracellular staining was observed through out the brain in both Tc1 and manage non transchromosomic mice. the original source Consequently, despite the fact that 9644 B may very well be a suita ble antibody for western blot studies of RRP1, it can’t be employed to determine Hsa21 favourable cells during the brains of Tc1 mice. Affinity purified antibody raised against RRP1 peptide B purified in the second rabbit did not recognise a Tc1 distinct band. A 50 kDa protein was weakly detected working with this antibody in sam ples of Tc1 and handle mouse brain, however, peptide B does not share any homology with mouse RRP1 therefore the 50 kDa band detected after probing with this antibody is extremely unlikely to get RRP1.
An antibody affinity purified against RRP1 peptide A did recognise a band constant together with the mole cular fat of RRP1 in samples of both Tc1 and con trol brain. Five of the nineteen amino acids of peptide A are homologous together with the mouse RRP1 pro tein sequence together with a sequence with large predicted antigenicity. screening compounds For that reason the antibody purified against peptide A may perhaps recognise each mouse and human RRP1 and consequently is not helpful to determine Hsa21 positive cells in the Tc1 model. An antibody affi nity purified towards peptide A through the other rabbit did not constantly recognise a band corre sponding to your molecular excess weight of RRP1. This suggests that RRP1 peptide A will not be a dependable anti gen for that production of rabbit polyclonal antibodies.
Antibodies that did not recognise a Tc1 unique product SOD1 Immunisation which has a single SOD1 peptide produced anti SOD1 antibodies that recognised a Tc1 particular band on western blots of complete brain protein. The size on the bands recognised is steady with all the regarded molecular weight from the SOD1 monomer. These antibodies also detected a band of the comparable molecular bodyweight in samples of complete brain proteins isolated from transgenic mice that more than express wild style or mutant human SOD1 and in samples of recombinant human SOD1. The 16 kDa band was not observed in samples of brain from non transchromosomic management mice. Nonetheless, soon after prolonged exposures a weak band that was smaller compared to the predominant 16 kDa band was detected by each 9637 and 9638 in Tc1 and manage mouse brain samples.
This smaller band could be mouse SOD1, therefore antibody 9637 and 9638 may weakly cross react with mouse SOD1. Moreover, these antibodies created an intracellular staining pattern of comparable intensity on Tc1 and non transchromosomic management mice brain sections, which were either paraffin embedded or cryopreserved. The antibody will not recognise cells specifi cally within the Tc1 brain and thus can’t be made use of to determine these Hsa21 constructive cells in our mouse model for future scientific studies. This outcome might take place mainly because the polyclonal antibodies produced recognise non SOD1 proteins and weakly cross react with mouse SOD1 in each Tc1 and control brain, or the antibodies produced only recognise denatured human SOD1.