Details of experiments between  Q15418  and Q5T2W1  Note that the results of all experiments are listed, regardless of the modification states of the fragments.
   Experiment series 1 Protein A Protein: RPS6KA1  Fragment: 725-735  Site: PBM  Sequence: RVRKLPSTTL  Construct: biotin-ttds-RRVRKLPSTTL 
Protein B Protein: NHERF3  Fragment: 2-107  Site: PDZ_1  Construct: his6-MBP-TEVsite-PDZ 
Average holdup BI:  0.42 
Holdup BI standard deviation:  0.03 
Immobilized partner concentration (10-6 M):  31 
Experiment method:  CALIP_holdup 
Details of affinity fitting:  BSA and lysozyme internal standards + bacterial cell lysate + normal layout 
Number of measurements:  4 
Measured pK d : 4.4 
    Experiment series 2 Protein A Protein: RPS6KA1  Fragment: 726-735  Site: PBM  Sequence: RVRKLPSTTL  Construct: Biotin-ado-ado-RVRKLPSTTL 
Protein B Protein: NHERF3  Fragment: 2-107  Site: PDZ_1  Construct: his6-MBP-TEVsite-PDZ 
Average holdup BI:  0.18 
Normalized holdup BI:  0.19 
Immobilized partner concentration (10-6 M):  18 
Experiment method:  DAPF_holdup 
Details of affinity fitting:  fluorescein + mCherry internal standards + normalization based on CALIP_holdup data + reverse layout 
Number of measurements:  1 
Measured pK d : 4.14 
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
   Experiment series 3 Protein A Protein: RPS6KA1  Fragment: 729-735  Site: PBM  Sequence: RVRKLPSTTL  Construct: Fluorescein-KLPSTTL 
Protein B Protein: NHERF3  Fragment: 2-107  Site: PDZ_1  Construct: his6-MBP-TEVsite-PDZ 
Measured dissociation constant (10-6 M):  80 
Standard deviation of dissociation constant (10-6 M):  20 
Experiment method:  FP_direct 
Number of measurements:  3 
Measured pK d : 4.1 
   Experiment series 4 Protein A Protein: RPS6KA1  Fragment: 725-735  Site: PBM  Sequence: RVRKLPSTTL  Construct: biotin-ttds-RRVRKLPSTTL 
Protein B Protein: NHERF3  Fragment: 2-107  Site: PDZ_1  Construct: his6-MBP-TEVsite-PDZ 
Average holdup BI:  0.15 
Holdup BI standard deviation:  0.01 
Normalized holdup BI:  0.16 
Normalized holdup BI standard deviation:  0.01 
Immobilized partner concentration (10-6 M):  18 
Experiment method:  DAPF_holdup 
Details of affinity fitting:  fluorescein + mCherry internal standards + normalization based on CALIP_holdup data + reverse layout 
Number of measurements:  2 
Measured pK d : 4.03 
   Experiment series 5 Protein A Protein: RPS6KA1  Fragment: 725-735  Site: PBM  Sequence: RVRKLPSTTL  Construct: biotin-ttds-RRVRKLPSTTL 
Protein B Protein: NHERF3  Fragment: 375-459  Site: PDZ_4  Construct: his6-MBP-TEVsite-PDZ 
Average holdup BI:  0.07 
Normalized holdup BI:  0.07 
Immobilized partner concentration (10-6 M):  18 
Experiment method:  DAPF_holdup 
Details of affinity fitting:  fluorescein + mCherry internal standards + normalization based on CALIP_holdup data + reverse layout 
Number of measurements:  1 
Measured pK d : 3.65 
   Experiment series 6 Protein A Protein: RPS6KA1  Fragment: 725-735  Site: PBM  Sequence: RVRKLPSTTL  Construct: biotin-ttds-RRVRKLPSTTL 
Protein B Protein: NHERF3  Fragment: 238-329  Site: PDZ_3  Construct: his6-MBP-TEVsite-PDZ 
Average holdup BI:  0.07 
Normalized holdup BI:  0.07 
Immobilized partner concentration (10-6 M):  18 
Experiment method:  DAPF_holdup 
Details of affinity fitting:  fluorescein + mCherry internal standards + normalization based on CALIP_holdup data + reverse layout 
Number of measurements:  1 
Measured pK d : 3.61 
   Experiment series 7 Protein A Protein: RPS6KA1  Fragment: 725-735  Site: PBM  Sequence: RVRKLPSTTL  Construct: biotin-ttds-RRVRKLPSTTL 
Protein B Protein: NHERF3  Fragment: 132-224  Site: PDZ_2  Construct: his6-MBP-TEVsite-PDZ 
Average holdup BI:  -0.03 
Holdup BI standard deviation:  0.02 
Immobilized partner concentration (10-6 M):  31 
Experiment method:  CALIP_holdup 
Details of affinity fitting:  BSA and lysozyme internal standards + bacterial cell lysate + normal layout 
Number of measurements:  4 
The affinity was below the detection threshold of the assay. 
   Experiment series 8 Protein A Protein: RPS6KA1  Fragment: 725-735  Site: PBM  Sequence: RVRKLPSTTL  Construct: Biotin-ttds-RRVRKLPSTTL 
Protein B Protein: NHERF3  Fragment: 2-107  Site: PDZ_1  Construct: his6-MBP-TEVsite-PDZ 
Measured dissociation constant (10-6 M):  0 
Experiment method:  SPR 
Number of measurements:  1 
The affinity was below the detection threshold of the assay. 
   Experiment series 9 Protein A Protein: RPS6KA1  Fragment: 725-735  Site: PBM  Sequence: RVRKLPSTTL  Construct: biotin-ttds-RRVRKLPSTTL 
Protein B Protein: NHERF3  Fragment: 2-107  Site: PDZ_1  Construct: his6-MBP-TEVsite-PDZ 
Measured dissociation constant (10-6 M):  0 
Experiment method:  competitive FP 
Details of affinity fitting:  Competitive binding equation in ProFit 
Number of measurements:  3 
The affinity was below the detection threshold of the assay. 
   Experiment series 10 Protein A Protein: RPS6KA1  Fragment: 725-735  Site: PBM  Sequence: RVRKLPSTTL  Construct: biotin-ttds-RRVRKLPSTTL 
Protein B Protein: NHERF3  Fragment: 238-329  Site: PDZ_3  Construct: his6-MBP-TEVsite-PDZ 
Average holdup BI:  0 
Holdup BI standard deviation:  0.04 
Immobilized partner concentration (10-6 M):  31 
Experiment method:  CALIP_holdup 
Details of affinity fitting:  BSA and lysozyme internal standards + bacterial cell lysate + normal layout 
Number of measurements:  4 
The affinity was below the detection threshold of the assay. 
   Experiment series 11 Protein A Protein: RPS6KA1  Fragment: 725-735  Site: PBM  Sequence: RVRKLPSTTL  Construct: biotin-ttds-RRVRKLPSTTL 
Protein B Protein: NHERF3  Fragment: 375-459  Site: PDZ_4  Construct: his6-MBP-TEVsite-PDZ 
Average holdup BI:  -0.03 
Holdup BI standard deviation:  0.06 
Immobilized partner concentration (10-6 M):  31 
Experiment method:  CALIP_holdup 
Details of affinity fitting:  BSA and lysozyme internal standards + bacterial cell lysate + normal layout 
Number of measurements:  4 
The affinity was below the detection threshold of the assay. 
   Experiment series 12 Protein A Protein: RPS6KA1  Fragment: 726-735  Site: PBM  Sequence: RVRKLPSTTL  Construct: Biotin-ado-ado-RVRKLPSTTL 
Protein B Protein: NHERF3  Fragment: 238-329  Site: PDZ_3  Construct: his6-MBP-TEVsite-PDZ 
Average holdup BI:  -0.01 
Normalized holdup BI:  -0.01 
Immobilized partner concentration (10-6 M):  18 
Experiment method:  DAPF_holdup 
Details of affinity fitting:  fluorescein + mCherry internal standards + normalization based on CALIP_holdup data + reverse layout 
Number of measurements:  1 
The affinity was below the detection threshold of the assay. 
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
   Experiment series 13 Protein A Protein: RPS6KA1  Fragment: 726-735  Site: PBM  Sequence: RVRKLPSTTL  Construct: Biotin-ado-ado-RVRKLPSTTL 
Protein B Protein: NHERF3  Fragment: 375-459  Site: PDZ_4  Construct: his6-MBP-TEVsite-PDZ 
Average holdup BI:  0.02 
Normalized holdup BI:  0.02 
Immobilized partner concentration (10-6 M):  18 
Experiment method:  DAPF_holdup 
Details of affinity fitting:  fluorescein + mCherry internal standards + normalization based on CALIP_holdup data + reverse layout 
Number of measurements:  1 
The affinity was below the detection threshold of the assay. 
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
   Experiment series 14 Protein A Protein: RPS6KA1  Fragment: 725-735  Site: PBM  Modification: (SEP)732  Sequence: RVRKLPSTTL  Construct: biotin-ttds-RRVRKLPpSTTL 
Protein B Protein: NHERF3  Fragment: 2-107  Site: PDZ_1  Construct: his6-MBP-TEVsite-PDZ 
Average holdup BI:  0.1 
Normalized holdup BI:  0.11 
Immobilized partner concentration (10-6 M):  15 
Experiment method:  DAPF_holdup 
Details of affinity fitting:  fluorescein + mCherry internal standards + normalization based on CALIP_holdup data + reverse layout 
Number of measurements:  1 
Measured pK d : 3.92 
Experimental Details not reported internal standards of holdup layout #5
   Experiment series 15 Protein A Protein: RPS6KA1  Fragment: 725-735  Site: PBM  Modification: (SEP)732  Sequence: RVRKLPSTTL  Construct: biotin-ttds-RRVRKLPpSTTL 
Protein B Protein: NHERF3  Fragment: 375-459  Site: PDZ_4  Construct: his6-MBP-TEVsite-PDZ 
Average holdup BI:  0.07 
Normalized holdup BI:  0.07 
Immobilized partner concentration (10-6 M):  15 
Experiment method:  DAPF_holdup 
Details of affinity fitting:  fluorescein + mCherry internal standards + normalization based on CALIP_holdup data + reverse layout 
Number of measurements:  1 
Measured pK d : 3.7 
Experimental Details not reported internal standards of holdup layout #5
   Experiment series 16 Protein A Protein: RPS6KA1  Fragment: 729-735  Site: PBM  Modification: (SEP)732  Sequence: RVRKLPSTTL  Construct: Fluorescein-KLP(pS)TTL 
Protein B Protein: NHERF3  Fragment: 2-107  Site: PDZ_1  Construct: his6-MBP-TEVsite-PDZ 
Measured dissociation constant (10-6 M):  220 
Standard deviation of dissociation constant (10-6 M):  30 
Experiment method:  FP_direct 
Number of measurements:  3 
Measured pK d : 3.66 
   Experiment series 17 Protein A Protein: RPS6KA1  Fragment: 725-735  Site: PBM  Modification: (SEP)732  Sequence: RVRKLPSTTL  Construct: biotin-ttds-RRVRKLPpSTTL 
Protein B Protein: NHERF3  Fragment: 2-107  Site: PDZ_1  Construct: his6-MBP-TEVsite-PDZ 
Average holdup BI:  0.11 
Holdup BI standard deviation:  0.14 
Immobilized partner concentration (10-6 M):  40 
Experiment method:  CALIP_holdup 
Details of affinity fitting:  BSA and lysozyme internal standards + bacterial cell lysate + normal layout 
Number of measurements:  3 
Measured pK d : 3.49 
   Experiment series 18 Protein A Protein: RPS6KA1  Fragment: 725-735  Site: PBM  Modification: (SEP)732  Sequence: RVRKLPSTTL  Construct: biotin-ttds-RRVRKLPpSTTL 
Protein B Protein: NHERF3  Fragment: 132-224  Site: PDZ_2  Construct: his6-MBP-TEVsite-PDZ 
Average holdup BI:  0.01 
Holdup BI standard deviation:  0.03 
Immobilized partner concentration (10-6 M):  40 
Experiment method:  CALIP_holdup 
Details of affinity fitting:  BSA and lysozyme internal standards + bacterial cell lysate + normal layout 
Number of measurements:  3 
The affinity was below the detection threshold of the assay. 
   Experiment series 19 Protein A Protein: RPS6KA1  Fragment: 725-735  Site: PBM  Modification: (SEP)732  Sequence: RVRKLPSTTL  Construct: Biotin-ttds-RRVRKLP(pS)TTL 
Protein B Protein: NHERF3  Fragment: 2-107  Site: PDZ_1  Construct: his6-MBP-TEVsite-PDZ 
Measured dissociation constant (10-6 M):  0 
Experiment method:  SPR 
Number of measurements:  1 
The affinity was below the detection threshold of the assay. 
   Experiment series 20 Protein A Protein: RPS6KA1  Fragment: 725-735  Site: PBM  Modification: (SEP)732  Sequence: RVRKLPSTTL  Construct: biotin-ttds-RRVRKLPpSTTL 
Protein B Protein: NHERF3  Fragment: 2-107  Site: PDZ_1  Construct: his6-MBP-TEVsite-PDZ 
Average holdup BI:  -0.01 
Holdup BI standard deviation:  0.01 
Normalized holdup BI:  -0.01 
Normalized holdup BI standard deviation:  0.01 
Immobilized partner concentration (10-6 M):  15 
Experiment method:  DAPF_holdup 
Details of affinity fitting:  fluorescein + mCherry internal standards + normalization based on CALIP_holdup data + reverse layout 
Number of measurements:  2 
The affinity was below the detection threshold of the assay. 
   Experiment series 21 Protein A Protein: RPS6KA1  Fragment: 725-735  Site: PBM  Modification: (SEP)732  Sequence: RVRKLPSTTL  Construct: biotin-ttds-RRVRKLPpSTTL 
Protein B Protein: NHERF3  Fragment: 2-107  Site: PDZ_1  Construct: his6-MBP-TEVsite-PDZ 
Measured dissociation constant (10-6 M):  0 
Experiment method:  competitive FP 
Details of affinity fitting:  Competitive binding equation in ProFit 
Number of measurements:  3 
The affinity was below the detection threshold of the assay. 
   Experiment series 22 Protein A Protein: RPS6KA1  Fragment: 725-735  Site: PBM  Modification: (SEP)732  Sequence: RVRKLPSTTL  Construct: biotin-ttds-RRVRKLPpSTTL 
Protein B Protein: NHERF3  Fragment: 238-329  Site: PDZ_3  Construct: his6-MBP-TEVsite-PDZ 
Average holdup BI:  0.02 
Normalized holdup BI:  0.02 
Immobilized partner concentration (10-6 M):  15 
Experiment method:  DAPF_holdup 
Details of affinity fitting:  fluorescein + mCherry internal standards + normalization based on CALIP_holdup data + reverse layout 
Number of measurements:  1 
The affinity was below the detection threshold of the assay. 
   Experiment series 23 Protein A Protein: RPS6KA1  Fragment: 725-735  Site: PBM  Modification: (SEP)732  Sequence: RVRKLPSTTL  Construct: biotin-ttds-RRVRKLPpSTTL 
Protein B Protein: NHERF3  Fragment: 238-329  Site: PDZ_3  Construct: his6-MBP-TEVsite-PDZ 
Average holdup BI:  0.04 
Normalized holdup BI:  0.04 
Immobilized partner concentration (10-6 M):  15 
Experiment method:  DAPF_holdup 
Details of affinity fitting:  fluorescein + mCherry internal standards + normalization based on CALIP_holdup data + reverse layout 
Number of measurements:  1 
The affinity was below the detection threshold of the assay. 
Experimental Details not reported internal standards of holdup layout #5
   Experiment series 24 Protein A Protein: RPS6KA1  Fragment: 725-735  Site: PBM  Modification: (SEP)732  Sequence: RVRKLPSTTL  Construct: biotin-ttds-RRVRKLPpSTTL 
Protein B Protein: NHERF3  Fragment: 238-329  Site: PDZ_3  Construct: his6-MBP-TEVsite-PDZ 
Average holdup BI:  0.04 
Holdup BI standard deviation:  0.01 
Immobilized partner concentration (10-6 M):  40 
Experiment method:  CALIP_holdup 
Details of affinity fitting:  BSA and lysozyme internal standards + bacterial cell lysate + normal layout 
Number of measurements:  3 
The affinity was below the detection threshold of the assay. 
   Experiment series 25 Protein A Protein: RPS6KA1  Fragment: 725-735  Site: PBM  Modification: (SEP)732  Sequence: RVRKLPSTTL  Construct: biotin-ttds-RRVRKLPpSTTL 
Protein B Protein: NHERF3  Fragment: 375-459  Site: PDZ_4  Construct: his6-MBP-TEVsite-PDZ 
Average holdup BI:  0.01 
Normalized holdup BI:  0.01 
Immobilized partner concentration (10-6 M):  15 
Experiment method:  DAPF_holdup 
Details of affinity fitting:  fluorescein + mCherry internal standards + normalization based on CALIP_holdup data + reverse layout 
Number of measurements:  1 
The affinity was below the detection threshold of the assay. 
   Experiment series 26 Protein A Protein: RPS6KA1  Fragment: 725-735  Site: PBM  Modification: (SEP)732  Sequence: RVRKLPSTTL  Construct: biotin-ttds-RRVRKLPpSTTL 
Protein B Protein: NHERF3  Fragment: 375-459  Site: PDZ_4  Construct: his6-MBP-TEVsite-PDZ 
Average holdup BI:  0.03 
Holdup BI standard deviation:  0.04 
Immobilized partner concentration (10-6 M):  40 
Experiment method:  CALIP_holdup 
Details of affinity fitting:  BSA and lysozyme internal standards + bacterial cell lysate + normal layout 
Number of measurements:  3 
The affinity was below the detection threshold of the assay. 
   Experiment series 27 Protein A Protein: RPS6KA1  Fragment: 725-735  Site: PBM  Modification: (TPO)733  Sequence: RVRKLPSTTL  Construct: biotin-ttds-RRVRKLPSpTTL 
Protein B Protein: NHERF3  Fragment: 2-107  Site: PDZ_1  Construct: his6-MBP-TEVsite-PDZ 
Average holdup BI:  -0 
Holdup BI standard deviation:  0 
Normalized holdup BI:  -0 
Normalized holdup BI standard deviation:  0 
Immobilized partner concentration (10-6 M):  18 
Experiment method:  DAPF_holdup 
Details of affinity fitting:  fluorescein + mCherry internal standards + normalization based on CALIP_holdup data + reverse layout 
Number of measurements:  2 
The affinity was below the detection threshold of the assay. 
   Experiment series 28 Protein A Protein: RPS6KA1  Fragment: 725-735  Site: PBM  Modification: (TPO)733  Sequence: RVRKLPSTTL  Construct: biotin-ttds-RRVRKLPSpTTL 
Protein B Protein: NHERF3  Fragment: 2-107  Site: PDZ_1  Construct: his6-MBP-TEVsite-PDZ 
Measured dissociation constant (10-6 M):  0 
Experiment method:  competitive FP 
Details of affinity fitting:  Competitive binding equation in ProFit 
Number of measurements:  3 
The affinity was below the detection threshold of the assay. 
   Experiment series 29 Protein A Protein: RPS6KA1  Fragment: 725-735  Site: PBM  Modification: (TPO)733  Sequence: RVRKLPSTTL  Construct: biotin-ttds-RRVRKLPSpTTL 
Protein B Protein: NHERF3  Fragment: 238-329  Site: PDZ_3  Construct: his6-MBP-TEVsite-PDZ 
Average holdup BI:  0.02 
Normalized holdup BI:  0.02 
Immobilized partner concentration (10-6 M):  18 
Experiment method:  DAPF_holdup 
Details of affinity fitting:  fluorescein + mCherry internal standards + normalization based on CALIP_holdup data + reverse layout 
Number of measurements:  1 
The affinity was below the detection threshold of the assay. 
   Experiment series 30 Protein A Protein: RPS6KA1  Fragment: 725-735  Site: PBM  Modification: (TPO)733  Sequence: RVRKLPSTTL  Construct: biotin-ttds-RRVRKLPSpTTL 
Protein B Protein: NHERF3  Fragment: 375-459  Site: PDZ_4  Construct: his6-MBP-TEVsite-PDZ 
Average holdup BI:  -0.01 
Normalized holdup BI:  -0.01 
Immobilized partner concentration (10-6 M):  18 
Experiment method:  DAPF_holdup 
Details of affinity fitting:  fluorescein + mCherry internal standards + normalization based on CALIP_holdup data + reverse layout 
Number of measurements:  1 
The affinity was below the detection threshold of the assay. 
   Experiment series 31 Protein A Protein: RPS6KA1  Fragment: 725-735  Site: PBM  Modification: (TPO)734  Sequence: RVRKLPSTTL  Construct: Biotin-ttds-RRVRKLPSTpTL  
Protein B Protein: NHERF3  Fragment: 2-107  Site: PDZ_1  Construct: his6-MBP-TEVsite-PDZ 
Average holdup BI:  0.25 
Normalized holdup BI:  0.28 
Immobilized partner concentration (10-6 M):  25 
Experiment method:  DAPF_holdup 
Details of affinity fitting:  fluorescein + mCherry internal standards + normalization based on CALIP_holdup data + reverse layout 
Number of measurements:  1 
Measured pK d : 4.2 
   Experiment series 32 Protein A Protein: RPS6KA1  Fragment: 725-735  Site: PBM  Modification: (TPO)734  Sequence: RVRKLPSTTL  Construct: Biotin-ttds-RRVRKLPSTpTL  
Protein B Protein: NHERF3  Fragment: 375-459  Site: PDZ_4  Construct: his6-MBP-TEVsite-PDZ 
Average holdup BI:  0.12 
Normalized holdup BI:  0.12 
Immobilized partner concentration (10-6 M):  25 
Experiment method:  DAPF_holdup 
Details of affinity fitting:  fluorescein + mCherry internal standards + normalization based on CALIP_holdup data + reverse layout 
Number of measurements:  1 
Measured pK d : 3.75 
   Experiment series 33 Protein A Protein: RPS6KA1  Fragment: 725-735  Site: PBM  Modification: (TPO)734  Sequence: RVRKLPSTTL  Construct: Biotin-ttds-RRVRKLPSTpTL  
Protein B Protein: NHERF3  Fragment: 238-329  Site: PDZ_3  Construct: his6-MBP-TEVsite-PDZ 
Average holdup BI:  0.05 
Normalized holdup BI:  0.05 
Immobilized partner concentration (10-6 M):  25 
Experiment method:  DAPF_holdup 
Details of affinity fitting:  fluorescein + mCherry internal standards + normalization based on CALIP_holdup data + reverse layout 
Number of measurements:  1 
Measured pK d : 3.36 
   Experiment series 34 Protein A Protein: RPS6KA1  Fragment: 725-735  Site: PBM  Modification: (TPO)734  Sequence: RVRKLPSTTL  Construct: Biotin-ttds-RRVRKLPSTpTL  
Protein B Protein: NHERF3  Fragment: 2-107  Site: PDZ_1  Construct: his6-MBP-TEVsite-PDZ 
Measured dissociation constant (10-6 M):  0 
Experiment method:  competitive FP 
Details of affinity fitting:  Competitive binding equation in ProFit 
Number of measurements:  3 
The affinity was below the detection threshold of the assay. 
    
 
Please cite ProfAff! (Gogl, G. et al., Nature Communications 2022)  
© The ProfAff database is developped by the research team of Gergo Gogl .