Details of experiments between Q14160 and P27228
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: 718-814
Protein A site: PDZ_1
Protein A construct: his6-MBP-TEVsite-PDZ

Protein B protein: HPV35-E6
Protein B fragment: 140-149
Protein B site: PBM
Protein B sequence: SKPTRRETEV
Protein B construct: biotin-ttds-SKPTRRETEV

Average holdup BI: 0.86
Immobilized partner concentration in holdup experiment (10-6M): 39.7
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: 5.24


Experiment series 2

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

Protein B protein: HPV35-E6
Protein B fragment: 140-149
Protein B site: PBM
Protein B sequence: SKPTRRETEV
Protein B construct: biotin-ttds-SKPTRRETEV

Average holdup BI: 0.78
Immobilized partner concentration in holdup experiment (10-6M): 39.7
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


Experiment series 3

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

Protein B protein: HPV35-E6
Protein B fragment: 140-149
Protein B site: PBM
Protein B sequence: SKPTRRETEV
Protein B construct: biotin-ttds-SKPTRRETEV

Average holdup BI: 0.67
Immobilized partner concentration in holdup experiment (10-6M): 39.7
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: 4.75


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: HPV35-E6
Protein B fragment: 140-149
Protein B site: PBM
Protein B sequence: SKPTRRETEV
Protein B construct: biotin-ttds-SKPTRRETEV

Average holdup BI: 0.19
Immobilized partner concentration in holdup experiment (10-6M): 39.7
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: 3.78


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: HPV35-E6
Protein B fragment: 140-149
Protein B site: PBM
Protein B sequence: SKPTRRETEV
Protein B construct: biotin-ttds-SKPTRRETEV

Measured dissociation constant (10-6M): 9.62
Standard deviation of dissociation constant (10-6M): 0.42
Experiment method: competitive FP
Details of affinity fitting: Competitive binding equation in ProFit

PUBMED ID of publication: 36115835
Number of measurements: 3
Measured pKd: 5.02


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: HPV35-E6
Protein B fragment: 140-149
Protein B site: PBM
Protein B sequence: SKPTRRETEV
Protein B construct: biotin-ttds-SKPTRRETEV

Measured dissociation constant (10-6M): 17.97
Standard deviation of dissociation constant (10-6M): 1.01
Experiment method: competitive FP
Details of affinity fitting: Competitive binding equation in ProFit

PUBMED ID of publication: 36115835
Number of measurements: 3
Measured pKd: 4.75


Experiment series 7

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

Protein B protein: HPV35-E6
Protein B fragment: 140-149
Protein B site: PBM
Protein B modification: (TPO)147
Protein B sequence: SKPTRRETEV
Protein B construct: Biotin-ttds-SKPTRREpTEV

Average holdup BI: -0.01
Normalized holdup BI: -0.01
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
The affinity was below the detection threshold of the assay.


Experiment series 8

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

Protein B protein: HPV35-E6
Protein B fragment: 140-149
Protein B site: PBM
Protein B modification: (TPO)147
Protein B sequence: SKPTRRETEV
Protein B construct: Biotin-ttds-SKPTRREpTEV

Average holdup BI: -0
Normalized holdup BI: -0
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
The affinity was below the detection threshold of the assay.


Experiment series 9

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

Protein B protein: HPV35-E6
Protein B fragment: 140-149
Protein B site: PBM
Protein B modification: (TPO)147
Protein B sequence: SKPTRRETEV
Protein B construct: Biotin-ttds-SKPTRREpTEV

Average holdup BI: 0.01
Normalized holdup BI: 0.01
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
The affinity was below the detection threshold of the assay.


Experiment series 10

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

Protein B protein: HPV35-E6
Protein B fragment: 140-149
Protein B site: PBM
Protein B sequence: SKPTRRETEV
Protein B construct: biotin-ttds-SKPTRRETEV

Average holdup BI: 0.5
Holdup BI standard deviation: 0.02
Normalized holdup BI: 0.56
Normalized holdup BI standard deviation: 0.02
Immobilized partner concentration in holdup experiment (10-6M): 18.55
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: 4.89


Experiment series 11

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

Protein B protein: HPV35-E6
Protein B fragment: 140-149
Protein B site: PBM
Protein B sequence: SKPTRRETEV
Protein B construct: biotin-ttds-SKPTRRETEV

Average holdup BI: 0.37
Holdup BI standard deviation: 0.01
Normalized holdup BI: 0.46
Normalized holdup BI standard deviation: 0.02
Immobilized partner concentration in holdup experiment (10-6M): 18.55
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: 4.7


Experiment series 12

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

Protein B protein: HPV35-E6
Protein B fragment: 140-149
Protein B site: PBM
Protein B sequence: SKPTRRETEV
Protein B construct: biotin-ttds-SKPTRRETEV

Average holdup BI: 0.36
Holdup BI standard deviation: 0.04
Normalized holdup BI: 0.38
Normalized holdup BI standard deviation: 0.04
Immobilized partner concentration in holdup experiment (10-6M): 18.55
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: 4.56


Experiment series 13

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

Protein B protein: HPV35-E6
Protein B fragment: 140-149
Protein B site: PBM
Protein B sequence: SKPTRRETEV
Protein B construct: biotin-ttds-SKPTRRETEV

Average holdup BI: 0.24
Normalized holdup BI: 0.3
Immobilized partner concentration in holdup experiment (10-6M): 18.55
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: 4.4


Experiment series 14

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

Protein B protein: HPV35-E6
Protein B fragment: 140-149
Protein B site: PBM
Protein B sequence: SKPTRRETEV
Protein B construct: biotin-ttds-SKPTRRETEV

Average holdup BI: 0.32
Normalized holdup BI: 0.35
Immobilized partner concentration in holdup experiment (10-6M): 18.55
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: 4.5


Experiment series 15

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

Protein B protein: HPV35-E6
Protein B fragment: 140-149
Protein B site: PBM
Protein B sequence: SKPTRRETEV
Protein B construct: biotin-ttds-SKPTRRETEV

Average holdup BI: 0.35
Normalized holdup BI: 0.46
Immobilized partner concentration in holdup experiment (10-6M): 18.55
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: 4.7


Experiment series 16

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

Protein B protein: HPV35-E6
Protein B fragment: 140-149
Protein B site: PBM
Protein B sequence: SKPTRRETEV
Protein B construct: biotin-ttds-SKPTRRETEV

Average holdup BI: 0.33
Normalized holdup BI: 0.4
Immobilized partner concentration in holdup experiment (10-6M): 18.55
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: 4.6


Experiment series 17

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

Protein B protein: HPV35-E6
Protein B fragment: 140-149
Protein B site: PBM
Protein B sequence: SKPTRRETEV
Protein B construct: biotin-ttds-SKPTRRETEV

Average holdup BI: 0.5
Normalized holdup BI: 0.59
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: 4.97


Experiment series 18

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

Protein B protein: HPV35-E6
Protein B fragment: 140-149
Protein B site: PBM
Protein B sequence: SKPTRRETEV
Protein B construct: biotin-ttds-SKPTRRETEV

Average holdup BI: 0.5
Normalized holdup BI: 0.5
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: 4.8


Experiment series 19

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

Protein B protein: HPV35-E6
Protein B fragment: 140-149
Protein B site: PBM
Protein B sequence: SKPTRRETEV
Protein B construct: biotin-ttds-SKPTRRETEV

Average holdup BI: 0.5
Normalized holdup BI: 0.5
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: 4.8