Details of experiments between A4U7F2 and Q86UL8
Note that the results of all experiments are listed, regardless of the modification states of the fragments.



Experiment series 1

Protein A protein: HPV67-E6
Protein A fragment: 140-149
Protein A site: PBM
Protein A sequence: SRPQRTQTQV
Protein A construct: biotin-ttds-SRPQRTQTQV

Protein B protein: MAGI2
Protein B fragment: 410-522
Protein B site: PDZ_2
Protein B construct: his6-MBP-TEVsite-PDZ

Measured dissociation constant (10-6M): 1.89
Standard deviation of dissociation constant (10-6M): 0.13
Experiment method: competitive FP
Details of affinity fitting: Competitive binding equation in ProFit

PUBMED ID of publication: 36115835
Number of measurements: 3
Measured pKd: 5.72


Experiment series 2

Protein A protein: HPV67-E6
Protein A fragment: 140-149
Protein A site: PBM
Protein A sequence: SRPQRTQTQV
Protein A construct: biotin-ttds-SRPQRTQTQV

Protein B protein: MAGI2
Protein B fragment: 410-522
Protein B site: PDZ_2
Protein B construct: his6-MBP-TEVsite-PDZ

Average holdup BI: 0.65
Immobilized partner concentration in holdup experiment (10-6M): 10
Experiment method: SAPF_holdup
Details of affinity fitting: fluorescein + mCherry internal standards + normal layout
PUBMED ID of publication: 36115835
Number of measurements: 1
Measured pKd: 5.39


Experiment series 3

Protein A protein: HPV67-E6
Protein A fragment: 140-149
Protein A site: PBM
Protein A sequence: SRPQRTQTQV
Protein A construct: biotin-ttds-SRPQRTQTQV

Protein B protein: MAGI2
Protein B fragment: 1-102
Protein B site: PDZ_1
Protein B construct: his6-MBP-TEVsite-PDZ

Average holdup BI: -0.02
Immobilized partner concentration in holdup experiment (10-6M): 10
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 4

Protein A protein: HPV67-E6
Protein A fragment: 140-149
Protein A site: PBM
Protein A sequence: SRPQRTQTQV
Protein A construct: biotin-ttds-SRPQRTQTQV

Protein B protein: MAGI2
Protein B fragment: 600-682
Protein B site: PDZ_3
Protein B construct: his6-MBP-TEVsite-PDZ

Average holdup BI: 0.04
Immobilized partner concentration in holdup experiment (10-6M): 10
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 5

Protein A protein: HPV67-E6
Protein A fragment: 140-149
Protein A site: PBM
Protein A sequence: SRPQRTQTQV
Protein A construct: biotin-ttds-SRPQRTQTQV

Protein B protein: MAGI2
Protein B fragment: 772-865
Protein B site: PDZ_4
Protein B construct: his6-MBP-TEVsite-PDZ

Average holdup BI: 0.08
Immobilized partner concentration in holdup experiment (10-6M): 10
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 6

Protein A protein: HPV67-E6
Protein A fragment: 140-149
Protein A site: PBM
Protein A sequence: SRPQRTQTQV
Protein A construct: biotin-ttds-SRPQRTQTQV

Protein B protein: MAGI2
Protein B fragment: 915-1017
Protein B site: PDZ_5
Protein B construct: his6-MBP-TEVsite-PDZ

Average holdup BI: 0.07
Immobilized partner concentration in holdup experiment (10-6M): 10
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 7

Protein A protein: HPV67-E6
Protein A fragment: 140-149
Protein A site: PBM
Protein A sequence: SRPQRTQTQV
Protein A construct: biotin-ttds-SRPQRTQTQV

Protein B protein: MAGI2
Protein B fragment: 1141-1229
Protein B site: PDZ_6
Protein B construct: his6-MBP-TEVsite-PDZ

Average holdup BI: -0.02
Immobilized partner concentration in holdup experiment (10-6M): 10
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 8

Protein A protein: HPV67-E6
Protein A fragment: 140-149
Protein A site: PBM
Protein A sequence: SRPQRTQTQV
Protein A construct: biotin-ttds-SRPQRTQTQV

Protein B protein: MAGI2
Protein B fragment: 410-522
Protein B site: PDZ_2
Protein B construct: his6-MBP-TEVsite-PDZ

Average holdup BI: 0.73
Normalized holdup BI: 0.8
Immobilized partner concentration in holdup experiment (10-6M): 22
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: 5.32


Experiment series 9

Protein A protein: HPV67-E6
Protein A fragment: 140-149
Protein A site: PBM
Protein A sequence: SRPQRTQTQV
Protein A construct: biotin-ttds-SRPQRTQTQV

Protein B protein: MAGI2
Protein B fragment: 772-865
Protein B site: PDZ_4
Protein B construct: his6-MBP-TEVsite-PDZ

Average holdup BI: 0.16
Normalized holdup BI: 0.22
Immobilized partner concentration in holdup experiment (10-6M): 22
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: 4.14


Experiment series 10

Protein A protein: HPV67-E6
Protein A fragment: 140-149
Protein A site: PBM
Protein A sequence: SRPQRTQTQV
Protein A construct: biotin-ttds-SRPQRTQTQV

Protein B protein: MAGI2
Protein B fragment: 915-1017
Protein B site: PDZ_5
Protein B construct: his6-MBP-TEVsite-PDZ

Average holdup BI: 0.16
Normalized holdup BI: 0.16
Immobilized partner concentration in holdup experiment (10-6M): 22
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: 3.94


Experiment series 11

Protein A protein: HPV67-E6
Protein A fragment: 140-149
Protein A site: PBM
Protein A sequence: SRPQRTQTQV
Protein A construct: biotin-ttds-SRPQRTQTQV

Protein B protein: MAGI2
Protein B fragment: 600-682
Protein B site: PDZ_3
Protein B construct: his6-MBP-TEVsite-PDZ

Average holdup BI: 0.04
Normalized holdup BI: 0.04
Immobilized partner concentration in holdup experiment (10-6M): 22
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 12

Protein A protein: HPV67-E6
Protein A fragment: 140-149
Protein A site: PBM
Protein A sequence: SRPQRTQTQV
Protein A construct: biotin-ttds-SRPQRTQTQV

Protein B protein: MAGI2
Protein B fragment: 1141-1229
Protein B site: PDZ_6
Protein B construct: his6-MBP-TEVsite-PDZ

Average holdup BI: 0.03
Normalized holdup BI: 0.04
Immobilized partner concentration in holdup experiment (10-6M): 22
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.