Info:(For Wildtype) Extinction Coefficient BGG: 0.0007 (y = 0.0007x Beer's Law Graph)Extinction Coefficient BSA: 0.0011 (y = 0.0011x Beer's Law Graph)Cell Lysate: ABS: 0.2, BSA Protein: 285.7mM, BGG Protein: 181.8mMCell Lysate 1 in 5 dilution: ABsorbtion, 0.03, BSA Protein: 214.3mM, BGG Protein: 136.4mMElution: Abs .03, Protein BSA: 42.9mM, Protein BGG: 27.3mMElution DIlution 1/5: ABS: 0.005, Protein BSA: 35.7mM, Protein BGG: 22.7mM Problem:5. Looking at the sequence of the BGG and BSA standards compared to the sequence of MBP-BAP, which ofthe two standards is the better choice for determining the concentration of MBP-BAP.A. What amino acids react with the Bradford reagent?B. Find the % Composition of amino acids that react with BradfordC. Which standard is best for determining the concentration of MBP-BAP? Briefly explain. D. Explain briefly why choosing a specific standard is less important for determining the concentration of proteinin the cell lysate compared to the elution fraction? For A I believe Cys, Tyr and Trp react with the Bradford reagent unsure how to solve rest of problem 5 however. Protein Sequences (from NCBI protein database, http://www.ncbi.nlm.nih.gov/genbank/)Bovine Serum Albumin (GenBank - NP_851335):DTHKSEIAHRFKDLGEEHFKGLVLIAFSQYLQQCPFDEHVKLVNELTE FAKTCVADESHAGCEKSLHTLFGDELCKVASLRETYGDMADCCEKQE PERNECFLSHKDDS PDLPKLKPDPNTLCDEFKADEKKFWGKYLYEIARRHPYFYAPELLYYANKYNGVFQECCQAEDKGACLLPKIETMREKVLASSARQRLRCASIQKFGERALKAWSVARLSQKFPKAEFVEVTKLVTDLTKVHKECCHGDLLECADDRADLAKYICDNQDTISSKLKECCDKPLLEKSHCIAEVEKDAI PENLP PLTADFAEDKDVCKNYQEAKDAFLGSFLYEYSRRHPEYAVSVLLRLAKEYEATLEECCAKDDPHACYSTVFDKLKHLVDEPONLIKQNCDQFEKLGEYGFQNALIVRYTRKVPQVSTPTLVEVSRSLGKVGTRCCTKPESERMPCTEDYLSLILNRLCVLHEKTPVSEKVTKCCTESLVNRRPCFSALTPDETYVPKAFDEKLFTFHADICTLPDTEKQIKKQTALVELLKHKPKATEEQLKTVMENFVAFVDKCCAADDKEACFAVEGPKLVVSTQTALAIg Gamma 1 C Chain (Heavy Chain of IgG; GenBank - P01857):note - bovine gamma globulin (BGG) is actually a mixture of different protein chains, with lg gamma 1 C chainbeing a prominent component of the mixtureASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTOKSLSLSPGKMBP-tagged Bacterial Alkaline PhosphataseMKIKTGARILALSALTTMMFSASALAKIEEGKLVIWINGDKGYNGLAEVGKKFEKDTGIKVTVEHPDKLEEKFPQVAATGDGPDIIFWAHDRFGGYAQSGLLAEITPDKAFQDKLYPFTWDAVRYNGKLIAYPIAVEALSLIYNKDLLPNPPKTWEEIPALDKELKAKGKSALMFNLQEPYFTWPLIAADGGYAFKYENGKYDIKDVGVDNAGAKAGLTFLVDLIKNKHMNADTDYSIAEAAFNKGETAMTINGPWAWSNIDTSKVNYGVTVLPTFKGQPSKPFVGVLSAGINAASPNKELAKEFLENYLLTDEGLEAVNKDKPLGAVALKSYEEELVKDPRIAATMENAQKGEIMPNIPOMSAFWYAVRTAVINAASGROTVDEALKDAQTNSSSNNNNNNNNNNLGIEGRTPEMPVLENRAAQGDI TAPGGARRLTGDQTAALRDSLSDKPAKNIILLIGDGMGDSEITAARNYAEGAGGFFKGIDALPLTGQYTHYALNKKTGKPDYVTDSAASATAWSTGVKTYNGALGVDIHEKDHPTILEMAKAAGLATGNVSTAELODATPAALVAHVTSRKCYGPSATSEKCPGNALEKGGKGSITEQLLNARADVTLGGGAKT FAETATAGEWQGKTLREQAQARGYQLVS DAASLNSVTEANQQKPL LGL FADGNMPVRWLGPKATYHGNIDKPAVTCTPNPQRNDSVPTLAQMTDKAIELLSKNEKGFFLQVEGASIDKQDHAANPCGQIGETVDLDEAVQRALEFAKKEGNTLVIVTADHAHASQIVAPDTKAPGLTQALNTKDGAVMVMSYGNSEEDSQEHTGSQLRIAAYGPHAANVVGLTDQTDLFYTMKAALGLK kDa25015010070502010Std. W.LadLys111W.W.Elu UBTM.LysM. Alk.Elu PhoBSA50BSA100ng/mL ng/mL

Question

Info:(For Wildtype) Extinction Coefficient BGG: 0.0007 (y = 0.0007x Beer's Law Graph)Extinction Coefficient BSA: 0.0011 (y = 0.0011x Beer's Law Graph)Cell Lysate: ABS: 0.2, BSA Protein: 285.7mM, BGG Protein: 181.8mMCell Lysate 1 in 5 dilution: ABsorbtion, 0.03, BSA Protein: 214.3mM, BGG Protein: 136.4mMElution: Abs .03, Protein BSA: 42.9mM, Protein BGG: 27.3mMElution DIlution 1/5: ABS: 0.005, Protein BSA: 35.7mM, Protein BGG: 22.7mM Problem:5. Looking at the sequence of the BGG and BSA standards compared to the sequence of MBP-BAP, which ofthe two standards is the better choice for determining the concentration of MBP-BAP.A. What amino acids react with the Bradford reagent?B. Find the % Composition of amino acids that react with BradfordC. Which standard is best for determining the concentration of MBP-BAP? Briefly explain. D. Explain briefly why choosing a specific standard is less important for determining the concentration of proteinin the cell lysate compared to the elution fraction? For A I believe Cys, Tyr and Trp react with the Bradford reagent unsure how to solve rest of problem 5 however. Protein Sequences (from NCBI protein database, http://www.ncbi.nlm.nih.gov/genbank/)Bovine Serum Albumin (GenBank - NP_851335):DTHKSEIAHRFKDLGEEHFKGLVLIAFSQYLQQCPFDEHVKLVNELTE FAKTCVADESHAGCEKSLHTLFGDELCKVASLRETYGDMADCCEKQE PERNECFLSHKDDS PDLPKLKPDPNTLCDEFKADEKKFWGKYLYEIARRHPYFYAPELLYYANKYNGVFQECCQAEDKGACLLPKIETMREKVLASSARQRLRCASIQKFGERALKAWSVARLSQKFPKAEFVEVTKLVTDLTKVHKECCHGDLLECADDRADLAKYICDNQDTISSKLKECCDKPLLEKSHCIAEVEKDAI PENLP PLTADFAEDKDVCKNYQEAKDAFLGSFLYEYSRRHPEYAVSVLLRLAKEYEATLEECCAKDDPHACYSTVFDKLKHLVDEPONLIKQNCDQFEKLGEYGFQNALIVRYTRKVPQVSTPTLVEVSRSLGKVGTRCCTKPESERMPCTEDYLSLILNRLCVLHEKTPVSEKVTKCCTESLVNRRPCFSALTPDETYVPKAFDEKLFTFHADICTLPDTEKQIKKQTALVELLKHKPKATEEQLKTVMENFVAFVDKCCAADDKEACFAVEGPKLVVSTQTALAIg Gamma 1 C Chain (Heavy Chain of IgG; GenBank - P01857):note - bovine gamma globulin (BGG) is actually a mixture of different protein chains, with lg gamma 1 C chainbeing a prominent component of the mixtureASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTOKSLSLSPGKMBP-tagged Bacterial Alkaline PhosphataseMKIKTGARILALSALTTMMFSASALAKIEEGKLVIWINGDKGYNGLAEVGKKFEKDTGIKVTVEHPDKLEEKFPQVAATGDGPDIIFWAHDRFGGYAQSGLLAEITPDKAFQDKLYPFTWDAVRYNGKLIAYPIAVEALSLIYNKDLLPNPPKTWEEIPALDKELKAKGKSALMFNLQEPYFTWPLIAADGGYAFKYENGKYDIKDVGVDNAGAKAGLTFLVDLIKNKHMNADTDYSIAEAAFNKGETAMTINGPWAWSNIDTSKVNYGVTVLPTFKGQPSKPFVGVLSAGINAASPNKELAKEFLENYLLTDEGLEAVNKDKPLGAVALKSYEEELVKDPRIAATMENAQKGEIMPNIPOMSAFWYAVRTAVINAASGROTVDEALKDAQTNSSSNNNNNNNNNNLGIEGRTPEMPVLENRAAQGDI TAPGGARRLTGDQTAALRDSLSDKPAKNIILLIGDGMGDSEITAARNYAEGAGGFFKGIDALPLTGQYTHYALNKKTGKPDYVTDSAASATAWSTGVKTYNGALGVDIHEKDHPTILEMAKAAGLATGNVSTAELODATPAALVAHVTSRKCYGPSATSEKCPGNALEKGGKGSITEQLLNARADVTLGGGAKT FAETATAGEWQGKTLREQAQARGYQLVS DAASLNSVTEANQQKPL LGL FADGNMPVRWLGPKATYHGNIDKPAVTCTPNPQRNDSVPTLAQMTDKAIELLSKNEKGFFLQVEGASIDKQDHAANPCGQIGETVDLDEAVQRALEFAKKEGNTLVIVTADHAHASQIVAPDTKAPGLTQALNTKDGAVMVMSYGNSEEDSQEHTGSQLRIAAYGPHAANVVGLTDQTDLFYTMKAALGLK kDa25015010070502010Std. W.LadLys111W.W.Elu UBTM.LysM. Alk.Elu PhoBSA50BSA100ng/mL ng/mL

Accepted Answer

In protein analysis and quantification, choosing the appropriate standard is crucial for accurate…