Details of experiments between the fragments Q5T2W1~238-329 and Q7Z628~587-596
Note that the results of all experiments are listed, regardless of the modification states of the fragments.



Experiment series 1

Protein A protein: NHERF3
Protein A fragment: 238-329
Protein A site: PDZ_3
Protein A construct: his6-MBP-TEVsite-PDZ

Protein B protein: NET1
Protein B fragment: 587-596
Protein B site: PBM
Protein B sequence: SGGKRKETLV
Protein B construct: biotin-ttds-SGGKRKETLV

Average holdup BI: 0.05
Immobilized partner concentration in holdup experiment (10-6M): 35
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
The affinity was below the detection threshold of the assay.


Experiment series 2

Protein A protein: NHERF3
Protein A fragment: 238-329
Protein A site: PDZ_3
Protein A construct: his6-MBP-TEVsite-PDZ

Protein B protein: NET1
Protein B fragment: 587-596
Protein B site: PBM
Protein B sequence: SGGKRKETLV
Protein B construct: biotin-ttds-SGGKRKETLV

Average holdup BI: 0
Normalized holdup BI: 0
Immobilized partner concentration in holdup experiment (10-6M): 17.9
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
The affinity was below the detection threshold of the assay.


Experiment series 3

Protein A protein: NHERF3
Protein A fragment: 238-329
Protein A site: PDZ_3
Protein A construct: his6-MBP-TEVsite-PDZ

Protein B protein: NET1
Protein B fragment: 587-596
Protein B site: PBM
Protein B sequence: SGGKRKETLV
Protein B construct: biotin-ttds-SGGKRKETLV

Average holdup BI: -0.03
Normalized holdup BI: -0.03
Immobilized partner concentration in holdup experiment (10-6M): 17.9
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
The affinity was below the detection threshold of the assay.


Experiment series 4

Protein A protein: NHERF3
Protein A fragment: 238-329
Protein A site: PDZ_3
Protein A construct: his6-MBP-TEVsite-PDZ

Protein B protein: NET1
Protein B fragment: 587-596
Protein B site: PBM
Protein B sequence: SGGKRKETLV
Protein B construct: Biotin-ado-ado-SGGKRKETLV

Average holdup BI: -0.03
Normalized holdup BI: -0.03
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: Crude peptide synthesis product. The peptide concentration in the affinity conversion step may differ from reality. Not reported internal standards of holdup layout #6
PUBMED ID of publication: 36115835
Number of measurements: 1
The affinity was below the detection threshold of the assay.