Details of experiments between Q96RT1 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: LAP2
Protein A fragment: 1314-1412
Protein A site: PDZ
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: 0.34
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: 4.12


Experiment series 2

Protein A protein: LAP2
Protein A fragment: 1314-1412
Protein A site: PDZ
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 3

Protein A protein: LAP2
Protein A fragment: 1314-1412
Protein A site: PDZ
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: 0.07
Holdup BI standard deviation: 0.02
Normalized holdup BI: 0.08
Normalized holdup BI standard deviation: 0.03
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: 4
Measured pKd: 3.69


Experiment series 4

Protein A protein: LAP2
Protein A fragment: 1314-1412
Protein A site: PDZ
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: 0.12
Normalized holdup BI: 0.13
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: 3.93


Experiment series 5

Protein A protein: LAP2
Protein A fragment: 1314-1412
Protein A site: PDZ
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: 0.07
Normalized holdup BI: 0.08
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: 3.68


Experiment series 6

Protein A protein: LAP2
Protein A fragment: 1314-1412
Protein A site: PDZ
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: 0.06
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: not reported internal standards of holdup layout #6
PUBMED ID of publication: 36115835
Number of measurements: 1
Measured pKd: 3.52