PLATE

The protein dissever from 70 per centime chroma of recrystallised ammonium sulphate was found to collect the maximum protein content (19.6 mg/g flower) and hence it was selected for merely studies and is abbreviated as PAF in the present study.4.2.1 Characterization of the selected PAF by Native rogue and SDS PAGE The selected PAF was characterized by column chromatography. This showed a single cap and was further characterized in native PAGE and SDS page. The results are shown in Plate the SDS-PAGE abstract of the ammonium sulphate precipitated protein extract showed 15 different protein flocks with good visibility in CBB R250 staining method (Figure).PLATE image IZATION OF PFPa BY PAGE AND SDS PAGEA Standard Protein MarkersB PAGE C SDS PAGE Each fraction showed a number of major and minor bands indicating several(prenominal) proteins. Of the several bands obtained in 70 per cent saturation of ammonium sulphate, the major band alone was eluted. In order to reclaim come f orward the presence of subunits in this band, it was further subjected to SDS PAGE.The results showed one major protein indicating the absence of subunits. The molecular weight of this protein was found to be 99 KD when compared to the standard molecular markers. 4.3. Fifty per centumage effective panelling of selected protein fraction of genus Plumeria alba The free fundamental scavenging capacity of perfluorocarbon was tested by its ability to decolourize the stable DPPH.The DPPH (2,2 diphenyl -1- picryl hydrazyl) substructure scavenging natural process was carried out using different concentrations of PFC (Figure). The radical scavenging activity was found to be superman dependent. Figure Percentage Free al-Qaida Scavenging Activity of Selected protein fraction of Plumeria alba The protein fraction of Plumeria flower extract showed the dose dependent DPPH radical scavenging activity.From the graph, the 50 percent effective concentrations of these were found to be 28 g a nd 35 g and employ in the further studies. Free radicals and their scavenging systems wanton away important role in the mend of normal and delayed types of wounds. The dose chemical reaction curve of DPPH radical scavenging activity 0 10 20 30 40 50 60 70 80 90 10010 20 30 40 50 60 70 80 90 100Ascorbic acid works sampleDPPH RadicalScavenging 9(%) of the extract and standards showed that at the highest concentration (0.5mg ml-1) the scavenging effect of the methanolic extract reached 9.3% (Afolayan et al., 2008). Shyuret al. (2005) also reported that the scavenging activity for free radicals of 1,1diphyryl-2-picrylhydrazyl (DPPH) has been widely utilise to assess the antioxidant activity of natural products from plants.The antioxidant activities of the leafy vegetables of India were careful in different systems of assays such as DPPH assay, super oxide radical scavenging assay, hydroxyl radical scavenging assay and lipid peroxidation assay and IC50 determine were calculated ( Dasgupta and De 2007).4.4. Matrix-Assisted Laser Desorption/Ionization judgment of conviction of Flight Mass Spectra (MALDI-TOF MS) MALDI-TOF mass spectra were used for the analysis of peptide mass fingerprinting and MS/MS ion search identification of the in-gel trypsin digested protein fragments (Figure 5), selected peptide populace were submitted to Mascot (http//www.matrixscience.com) for SwissProt databases search. There was no conclusive scar in peptide mass fingerprinting, since MS/MS ion search program was selected for further identification.The MS/MS ion search in the NCBIProt database revealed that, WRKY placement factor WRKY24-like isoform X1 Juglans regia (Figure 6-7) with the protein take a leak of 81 (Protein score is 10*Log(P), where P is the probability that the observe match is a random event. Protein gain ground greater than 80 are fundamental (p