Details of experiments between the fragments Q6ZMN7~214-318 and Q15311~646-655
Note that the results of all experiments are listed, regardless of the modification states of the fragments.
Experiment series 1
Protein A protein: PDZRN4
Protein A fragment: 214-318
Protein A site: PDZ_1
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: RALBP1
Protein B fragment: 646-655
Protein B site: PBM
Protein B sequence: PSRDRKETSI
Protein B construct: biotin-ttds-PSRDRKETSI
Average holdup BI: 0.05
Immobilized partner concentration in holdup experiment (10-6M): 20
Experiment method: SAPF_holdup
Details of affinity fitting: fluorescein + mCherry internal standards + normal layout
PUBMED ID of publication: 36115835
Number of measurements: 1
The affinity was below the detection threshold of the assay.
Experiment series 2
Protein A protein: PDZRN4
Protein A fragment: 214-318
Protein A site: PDZ_1
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: RALBP1
Protein B fragment: 646-655
Protein B site: PBM
Protein B sequence: PSRDRKETSI
Protein B construct: biotin-ttds-PSRDRKETSI
Average holdup BI: 0.49
Normalized holdup BI: 0.49
Immobilized partner concentration in holdup experiment (10-6M): 28
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: 4.57
Experiment series 3
Protein A protein: PDZRN4
Protein A fragment: 214-318
Protein A site: PDZ_1
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: RALBP1
Protein B fragment: 646-655
Protein B site: PBM
Protein B sequence: PSRDRKETSI
Protein B construct: biotin-ttds-PSRDRKETSI
Average holdup BI: 0.49
Normalized holdup BI: 0.54
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: 4.87