Details of experiments between O14924 and Q14160
Note that the results of all experiments are listed, regardless of the modification states of the fragments.



Experiment series 1

Protein A protein: RGS12
Protein A fragment: 1438-1447
Protein A site: PBM
Protein A sequence: KTSAHHATFV
Protein A construct: Biotin-Ahx-KTSAHHATFV

Protein B protein: SCRIB
Protein B fragment: 999-1093
Protein B site: PDZ_3
Protein B construct: his6-MBP-TEVsite-PDZ

Average holdup BI: 0.17
Normalized holdup BI: 0.21
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
PUBMED ID of publication: 36115835
Number of measurements: 1
Measured pKd: 4.18


Experiment series 2

Protein A protein: RGS12
Protein A fragment: 1438-1447
Protein A site: PBM
Protein A sequence: KTSAHHATFV
Protein A construct: Biotin-Ahx-KTSAHHATFV

Protein B protein: SCRIB
Protein B fragment: 718-814
Protein B site: PDZ_1
Protein B construct: his6-MBP-TEVsite-PDZ

Average holdup BI: 0.08
Holdup BI standard deviation: 0.02
Normalized holdup BI: 0.1
Normalized holdup BI standard deviation: 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
PUBMED ID of publication: 36115835
Number of measurements: 2
Measured pKd: 3.8


Experiment series 3

Protein A protein: RGS12
Protein A fragment: 1438-1447
Protein A site: PBM
Protein A sequence: KTSAHHATFV
Protein A construct: Biotin-Ahx-KTSAHHATFV

Protein B protein: SCRIB
Protein B fragment: 858-949
Protein B site: PDZ_2
Protein B construct: his6-MBP-TEVsite-PDZ

Average holdup BI: 0.04
Normalized holdup BI: 0.05
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
PUBMED ID of publication: 36115835
Number of measurements: 1
Measured pKd: 3.47


Experiment series 4

Protein A protein: RGS12
Protein A fragment: 1438-1447
Protein A site: PBM
Protein A sequence: KTSAHHATFV
Protein A construct: Biotin-ado-ado-KTSAHHATFV

Protein B protein: SCRIB
Protein B fragment: 999-1093
Protein B site: PDZ_3
Protein B construct: his6-MBP-TEVsite-PDZ

Average holdup BI: 0.28
Normalized holdup BI: 0.32
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
Measured pKd: 4.46


Experiment series 5

Protein A protein: RGS12
Protein A fragment: 1438-1447
Protein A site: PBM
Protein A sequence: KTSAHHATFV
Protein A construct: Biotin-ado-ado-KTSAHHATFV

Protein B protein: SCRIB
Protein B fragment: 718-814
Protein B site: PDZ_1
Protein B construct: his6-MBP-TEVsite-PDZ

Average holdup BI: 0.06
Normalized holdup BI: 0.06
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
Measured pKd: 3.52


Experiment series 6

Protein A protein: RGS12
Protein A fragment: 1438-1447
Protein A site: PBM
Protein A sequence: KTSAHHATFV
Protein A construct: Biotin-ado-ado-KTSAHHATFV

Protein B protein: SCRIB
Protein B fragment: 858-949
Protein B site: PDZ_2
Protein B construct: his6-MBP-TEVsite-PDZ

Average holdup BI: 0.08
Normalized holdup BI: 0.08
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
Measured pKd: 3.68