Details of experiments between the fragments Q6P0Q8~1102-1193 and P60484~393-403
Note that the results of all experiments are listed, regardless of the modification states of the fragments.
Experiment series 1
Protein A protein: MAST2
Protein A fragment: 1102-1193
Protein A site: PDZ
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: PTEN
Protein B fragment: 393-403
Protein B site: PBM
Protein B modification: (ALY)402
Protein B sequence: EDQHTQITKV
Protein B construct: biotin-ttds-DEDQHTQITKacV
Average holdup BI: 0.86
Immobilized partner concentration in holdup experiment (10-6M): 29
Experiment method: CALIP_holdup
Details of affinity fitting: BSA and lysozyme internal standards + bacterial cell lysate + normal layout
PUBMED ID of publication: 36115835
Number of measurements: 1
Measured pKd: 5.37
Experiment series 2
Protein A protein: MAST2
Protein A fragment: 1102-1193
Protein A site: PDZ
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: PTEN
Protein B fragment: 393-403
Protein B site: PBM
Protein B sequence: EDQHTQITKV
Protein B construct: biotin-ttds-DEDQHTQITKV
Average holdup BI: 0.71
Immobilized partner concentration in holdup experiment (10-6M): 36
Experiment method: CALIP_holdup
Details of affinity fitting: BSA and lysozyme internal standards + bacterial cell lysate + normal layout
PUBMED ID of publication: 36115835
Number of measurements: 1
Measured pKd: 4.87
Experiment series 3
Protein A protein: MAST2
Protein A fragment: 1102-1193
Protein A site: PDZ
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: PTEN
Protein B fragment: 393-403
Protein B site: PBM
Protein B sequence: EDQHTQITKV
Protein B construct: biotin-ttds-DEDQHTQITKV
Measured dissociation constant (10-6M): 3.14
Standard deviation of dissociation constant (10-6M): 0.18
Experiment method: competitive FP
Details of affinity fitting: Competitive binding equation in ProFit
PUBMED ID of publication: 36115835
Number of measurements: 3
Measured pKd: 5.5
Experiment series 4
Protein A protein: MAST2
Protein A fragment: 1102-1193
Protein A site: PDZ
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: PTEN
Protein B fragment: 393-403
Protein B site: PBM
Protein B modification: (ALY)402
Protein B sequence: EDQHTQITKV
Protein B construct: biotin-ttds-DEDQHTQITKacV
Measured dissociation constant (10-6M): 2.56
Standard deviation of dissociation constant (10-6M): 0.13
Experiment method: competitive FP
Details of affinity fitting: Competitive binding equation in ProFit
PUBMED ID of publication: 36115835
Number of measurements: 3
Measured pKd: 5.59
Experiment series 5
Protein A protein: MAST2
Protein A fragment: 1102-1193
Protein A site: PDZ
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: PTEN
Protein B fragment: 393-403
Protein B site: PBM
Protein B modification: (ALY)402
Protein B sequence: EDQHTQITKV
Protein B construct: biotin-ttds-DEDQHTQITKacV
Average holdup BI: 0.66
Normalized holdup BI: 0.66
Immobilized partner concentration in holdup experiment (10-6M): 17
Experiment method: DAPF_holdup
Details of affinity fitting: fluorescein + mCherry internal standards + normalization based on CALIP_holdup data + reverse layout
PUBMED ID of publication: 36115835
Number of measurements: 1
Measured pKd: 5.14
Experiment series 6
Protein A protein: MAST2
Protein A fragment: 1102-1193
Protein A site: PDZ
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: PTEN
Protein B fragment: 393-403
Protein B site: PBM
Protein B sequence: EDQHTQITKV
Protein B construct: biotin-ttds-DEDQHTQITKV
Average holdup BI: 0.74
Normalized holdup BI: 0.74
Immobilized partner concentration in holdup experiment (10-6M): 19
Experiment method: DAPF_holdup
Details of affinity fitting: fluorescein + mCherry internal standards + normalization based on CALIP_holdup data + reverse layout
PUBMED ID of publication: 36115835
Number of measurements: 1
Measured pKd: 5.24
Experiment series 7
Protein A protein: MAST2
Protein A fragment: 1102-1193
Protein A site: PDZ
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: PTEN
Protein B fragment: 393-403
Protein B site: PBM
Protein B modification: (ALY)402
Protein B sequence: EDQHTQITKV
Protein B construct: biotin-ttds-DEDQHTQITKacV
Average holdup BI: 0.77
Normalized holdup BI: 0.85
Immobilized partner concentration in holdup experiment (10-6M): 17
Experiment method: DAPF_holdup
Details of affinity fitting: fluorescein + mCherry internal standards + normalization based on CALIP_holdup data + reverse layout
Experimental details: not reported internal standards of holdup layout #5
Number of measurements: 1
Measured pKd: 5.61
Experiment series 8
Protein A protein: MAST2
Protein A fragment: 1102-1193
Protein A site: PDZ
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: PTEN
Protein B fragment: 393-403
Protein B site: PBM
Protein B modification: (ALY)402
Protein B sequence: EDQHTQITKV
Protein B construct: biotin-ttds-DEDQHTQITKacV
Average holdup BI: 0.82
Normalized holdup BI: 0.82
Immobilized partner concentration in holdup experiment (10-6M): 17
Experiment method: DAPF_holdup
Details of affinity fitting: fluorescein + mCherry internal standards + normalization based on CALIP_holdup data + reverse layout
Experimental details: not reported internal standards of holdup layout #5
Number of measurements: 1
Measured pKd: 5.53
Experiment series 9
Protein A protein: MAST2
Protein A fragment: 1102-1193
Protein A site: PDZ
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: PTEN
Protein B fragment: 393-403
Protein B site: PBM
Protein B modification: (ALY)402
Protein B sequence: EDQHTQITKV
Protein B construct: biotin-ttds-DEDQHTQITKacV
Average holdup BI: 0.89
Normalized holdup BI: 0.89
Immobilized partner concentration in holdup experiment (10-6M): 18
Experiment method: DAPF_holdup
Details of affinity fitting: fluorescein + mCherry internal standards + normalization based on CALIP_holdup data + reverse layout
Experimental details: not reported internal standards of holdup layout #6
PUBMED ID of publication: 36115835
Number of measurements: 1
Measured pKd: 5.77