Details of experiments between Q14160 and Q7Z628
Note that the results of all experiments are listed, regardless of the modification states of the fragments.



Experiment series 1

Protein A protein: SCRIB
Protein A fragment: 858-949
Protein A site: PDZ_2
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: 0.36
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
Measured pKd: 4.22


Experiment series 2

Protein A protein: SCRIB
Protein A fragment: 718-814
Protein A site: PDZ_1
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: 0.32
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
Measured pKd: 4.13


Experiment series 3

Protein A protein: SCRIB
Protein A fragment: 999-1093
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: 0.23
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
Measured pKd: 3.94


Experiment series 4

Protein A protein: SCRIB
Protein A fragment: 1090-1197
Protein A site: PDZ_4
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: -0.13
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 5

Protein A protein: SCRIB
Protein A fragment: 999-1093
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

Measured dissociation constant (10-6M): 43.01
Standard deviation of dissociation constant (10-6M): 7.18
Experiment method: competitive FP
Details of affinity fitting: Competitive binding equation in ProFit

PUBMED ID of publication: 36115835
Number of measurements: 3
Measured pKd: 4.37


Experiment series 6

Protein A protein: SCRIB
Protein A fragment: 718-814
Protein A site: PDZ_1
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

Measured dissociation constant (10-6M): 101.31
Standard deviation of dissociation constant (10-6M): 4.5
Experiment method: competitive FP
Details of affinity fitting: Competitive binding equation in ProFit

PUBMED ID of publication: 36115835
Number of measurements: 3
Measured pKd: 3.99


Experiment series 7

Protein A protein: SCRIB
Protein A fragment: 999-1093
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: 0.23
Holdup BI standard deviation: 0.12
Normalized holdup BI: 0.26
Normalized holdup BI standard deviation: 0.14
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: 2
Measured pKd: 4.31


Experiment series 8

Protein A protein: SCRIB
Protein A fragment: 718-814
Protein A site: PDZ_1
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: 0.14
Holdup BI standard deviation: 0.06
Normalized holdup BI: 0.15
Normalized holdup BI standard deviation: 0.06
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: 8
Measured pKd: 4


Experiment series 9

Protein A protein: SCRIB
Protein A fragment: 858-949
Protein A site: PDZ_2
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
Holdup BI standard deviation: 0.02
Normalized holdup BI: 0.06
Normalized holdup BI standard deviation: 0.02
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: 2
Measured pKd: 3.54


Experiment series 10

Protein A protein: SCRIB
Protein A fragment: 718-814
Protein A site: PDZ_1
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: 0.02
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 11

Protein A protein: SCRIB
Protein A fragment: 718-814
Protein A site: PDZ_1
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: 0.06
Normalized holdup BI: 0.07
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
Measured pKd: 3.63


Experiment series 12

Protein A protein: SCRIB
Protein A fragment: 858-949
Protein A site: PDZ_2
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: 0.07
Normalized holdup BI: 0.09
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
Measured pKd: 3.73


Experiment series 13

Protein A protein: SCRIB
Protein A fragment: 999-1093
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: 0.13
Normalized holdup BI: 0.16
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
Measured pKd: 4.03


Experiment series 14

Protein A protein: SCRIB
Protein A fragment: 999-1093
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: 0.26
Normalized holdup BI: 0.3
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: 4.41


Experiment series 15

Protein A protein: SCRIB
Protein A fragment: 718-814
Protein A site: PDZ_1
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: 0.21
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
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: 4.2


Experiment series 16

Protein A protein: SCRIB
Protein A fragment: 858-949
Protein A site: PDZ_2
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: 0.19
Normalized holdup BI: 0.19
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: 4.12