Details of experiments between  Q12959  and Q15418  Note that the results of all experiments are listed, regardless of the modification states of the fragments.
   Experiment series 1 Protein A Protein: DLG1  Fragment: 314-412  Site: PDZ_2  Construct: his6-MBP-TEVsite-PDZ 
Protein B Protein: RPS6KA1  Fragment: 725-735  Site: PBM  Sequence: RVRKLPSTTL  Construct: biotin-ttds-RRVRKLPSTTL 
Average holdup BI:  0.07 
Normalized holdup BI:  0.08 
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 : 3.7 
    Experiment series 2 Protein A Protein: DLG1  Fragment: 457-570  Site: PDZ_3  Construct: his6-MBP-TEVsite-PDZ 
Protein B Protein: RPS6KA1  Fragment: 725-735  Site: PBM  Sequence: RVRKLPSTTL  Construct: biotin-ttds-RRVRKLPSTTL 
Average holdup BI:  0.06 
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 : 3.63 
   Experiment series 3 Protein A Protein: DLG1  Fragment: 221-314  Site: PDZ_1  Construct: avi(biotin)-his6-MBP-TEVsite-PDZ 
Protein B Protein: RPS6KA1  Fragment: 1-735  Construct: endogenous protein (Jurkat) 
Average holdup BI:  0.01 
Immobilized partner concentration (10-6 M):  10 
P value:  0.16 
Experiment method:  nHU_Jurkat 
Details of affinity fitting:  hyperbolic binding equation 
Date of measurement:  2022-08-03 
Number of measurements:  3 
The affinity was below the detection threshold of the assay. 
Experimental Details -log10(P Two-sided unpaired T-test (6 vs 3))
   Experiment series 4 Protein A Protein: DLG1  Fragment: 221-314  Site: PDZ_1  Construct: his6-MBP-TEVsite-PDZ 
Protein B Protein: RPS6KA1  Fragment: 725-735  Site: PBM  Sequence: RVRKLPSTTL  Construct: biotin-ttds-RRVRKLPSTTL 
Average holdup BI:  0.03 
Normalized holdup BI:  0.04 
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 5 Protein A Protein: DLG1  Fragment: 221-314  Site: PDZ_1  Construct: his6-MBP-TEVsite-PDZ 
Protein B Protein: RPS6KA1  Fragment: 725-735  Site: PBM  Sequence: RVRKLPSTTL  Construct: biotin-ttds-RRVRKLPSTTL 
Average holdup BI:  0.06 
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 
The affinity was below the detection threshold of the assay. 
   Experiment series 6 Protein A Protein: DLG1  Fragment: 221-314  Site: PDZ_1  Construct: his6-MBP-TEVsite-PDZ 
Protein B Protein: RPS6KA1  Fragment: 726-735  Site: PBM  Sequence: RVRKLPSTTL  Construct: Biotin-ado-ado-RVRKLPSTTL 
Average holdup BI:  0.04 
Normalized holdup BI:  0.05 
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 7 Protein A Protein: DLG1  Fragment: 314-412  Site: PDZ_2  Construct: avi(biotin)-his6-MBP-TEVsite-PDZ 
Protein B Protein: RPS6KA1  Fragment: 1-735  Construct: endogenous protein (Jurkat) 
Average holdup BI:  0.02 
Immobilized partner concentration (10-6 M):  10 
P value:  0.42 
Experiment method:  nHU_Jurkat 
Details of affinity fitting:  hyperbolic binding equation 
Date of measurement:  2022-08-03 
Number of measurements:  3 
The affinity was below the detection threshold of the assay. 
Experimental Details -log10(P Two-sided unpaired T-test (6 vs 3))
   Experiment series 8 Protein A Protein: DLG1  Fragment: 314-412  Site: PDZ_2  Construct: his6-MBP-TEVsite-PDZ 
Protein B Protein: RPS6KA1  Fragment: 725-735  Site: PBM  Sequence: RVRKLPSTTL  Construct: biotin-ttds-RRVRKLPSTTL 
Average holdup BI:  0.06 
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 
The affinity was below the detection threshold of the assay. 
   Experiment series 9 Protein A Protein: DLG1  Fragment: 314-412  Site: PDZ_2  Construct: his6-MBP-TEVsite-PDZ 
Protein B Protein: RPS6KA1  Fragment: 726-735  Site: PBM  Sequence: RVRKLPSTTL  Construct: Biotin-ado-ado-RVRKLPSTTL 
Average holdup BI:  0.04 
Normalized holdup BI:  0.04 
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 10 Protein A Protein: DLG1  Fragment: 457-570  Site: PDZ_3  Construct: his6-MBP-TEVsite-PDZ 
Protein B Protein: RPS6KA1  Fragment: 725-735  Site: PBM  Sequence: RVRKLPSTTL  Construct: biotin-ttds-RRVRKLPSTTL 
Average holdup BI:  0.08 
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: DLG1  Fragment: 457-570  Site: PDZ_3  Construct: his6-MBP-TEVsite-PDZ 
Protein B Protein: RPS6KA1  Fragment: 726-735  Site: PBM  Sequence: RVRKLPSTTL  Construct: Biotin-ado-ado-RVRKLPSTTL 
Average holdup BI:  -0.05 
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 
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 12 Protein A Protein: DLG1  Fragment: 314-412  Site: PDZ_2  Construct: his6-MBP-TEVsite-PDZ 
Protein B Protein: RPS6KA1  Fragment: 725-735  Site: PBM  Modification: (SEP)732  Sequence: RVRKLPSTTL  Construct: biotin-ttds-RRVRKLPpSTTL 
Average holdup BI:  0.08 
Normalized holdup BI:  0.09 
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 : 3.84 
Experimental Details not reported internal standards of holdup layout #5
   Experiment series 13 Protein A Protein: DLG1  Fragment: 457-570  Site: PDZ_3  Construct: his6-MBP-TEVsite-PDZ 
Protein B Protein: RPS6KA1  Fragment: 725-735  Site: PBM  Modification: (SEP)732  Sequence: RVRKLPSTTL  Construct: biotin-ttds-RRVRKLPpSTTL 
Average holdup BI:  0.06 
Normalized holdup BI:  0.08 
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 : 3.8 
Experimental Details not reported internal standards of holdup layout #5
   Experiment series 14 Protein A Protein: DLG1  Fragment: 221-314  Site: PDZ_1  Construct: his6-MBP-TEVsite-PDZ 
Protein B Protein: RPS6KA1  Fragment: 725-735  Site: PBM  Modification: (SEP)732  Sequence: RVRKLPSTTL  Construct: biotin-ttds-RRVRKLPpSTTL 
Average holdup BI:  0.06 
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 : 3.73 
Experimental Details not reported internal standards of holdup layout #5
   Experiment series 15 Protein A Protein: DLG1  Fragment: 221-314  Site: PDZ_1  Construct: his6-MBP-TEVsite-PDZ 
Protein B Protein: RPS6KA1  Fragment: 725-735  Site: PBM  Modification: (SEP)732  Sequence: RVRKLPSTTL  Construct: biotin-ttds-RRVRKLPpSTTL 
Average holdup BI:  0.06 
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 : 3.69 
   Experiment series 16 Protein A Protein: DLG1  Fragment: 314-412  Site: PDZ_2  Construct: his6-MBP-TEVsite-PDZ 
Protein B Protein: RPS6KA1  Fragment: 725-735  Site: PBM  Modification: (SEP)732  Sequence: RVRKLPSTTL  Construct: biotin-ttds-RRVRKLPpSTTL 
Average holdup BI:  0.05 
Normalized holdup BI:  0.06 
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 : 3.61 
   Experiment series 17 Protein A Protein: DLG1  Fragment: 221-314  Site: PDZ_1  Construct: his6-MBP-TEVsite-PDZ 
Protein B Protein: RPS6KA1  Fragment: 725-735  Site: PBM  Modification: (SEP)732  Sequence: RVRKLPSTTL  Construct: biotin-ttds-RRVRKLPpSTTL 
Average holdup BI:  0.03 
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 18 Protein A Protein: DLG1  Fragment: 314-412  Site: PDZ_2  Construct: his6-MBP-TEVsite-PDZ 
Protein B Protein: RPS6KA1  Fragment: 725-735  Site: PBM  Modification: (SEP)732  Sequence: RVRKLPSTTL  Construct: biotin-ttds-RRVRKLPpSTTL 
Average holdup BI:  0.05 
Holdup BI standard deviation:  0.02 
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: DLG1  Fragment: 457-570  Site: PDZ_3  Construct: his6-MBP-TEVsite-PDZ 
Protein B Protein: RPS6KA1  Fragment: 725-735  Site: PBM  Modification: (SEP)732  Sequence: RVRKLPSTTL  Construct: biotin-ttds-RRVRKLPpSTTL 
Average holdup BI:  -0.03 
Normalized holdup BI:  -0.03 
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 20 Protein A Protein: DLG1  Fragment: 457-570  Site: PDZ_3  Construct: his6-MBP-TEVsite-PDZ 
Protein B Protein: RPS6KA1  Fragment: 725-735  Site: PBM  Modification: (SEP)732  Sequence: RVRKLPSTTL  Construct: biotin-ttds-RRVRKLPpSTTL 
Average holdup BI:  0 
Holdup BI standard deviation:  0.07 
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 21 Protein A Protein: DLG1  Fragment: 221-314  Site: PDZ_1  Construct: his6-MBP-TEVsite-PDZ 
Protein B Protein: RPS6KA1  Fragment: 725-735  Site: PBM  Modification: (TPO)733  Sequence: RVRKLPSTTL  Construct: biotin-ttds-RRVRKLPSpTTL 
Average holdup BI:  0.05 
Normalized holdup BI:  0.06 
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 : 3.58 
   Experiment series 22 Protein A Protein: DLG1  Fragment: 314-412  Site: PDZ_2  Construct: his6-MBP-TEVsite-PDZ 
Protein B Protein: RPS6KA1  Fragment: 725-735  Site: PBM  Modification: (TPO)733  Sequence: RVRKLPSTTL  Construct: biotin-ttds-RRVRKLPSpTTL 
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 23 Protein A Protein: DLG1  Fragment: 457-570  Site: PDZ_3  Construct: his6-MBP-TEVsite-PDZ 
Protein B Protein: RPS6KA1  Fragment: 725-735  Site: PBM  Modification: (TPO)733  Sequence: RVRKLPSTTL  Construct: biotin-ttds-RRVRKLPSpTTL 
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 24 Protein A Protein: DLG1  Fragment: 314-412  Site: PDZ_2  Construct: his6-MBP-TEVsite-PDZ 
Protein B Protein: RPS6KA1  Fragment: 725-735  Site: PBM  Modification: (TPO)734  Sequence: RVRKLPSTTL  Construct: Biotin-ttds-RRVRKLPSTpTL  
Average holdup BI:  0.12 
Normalized holdup BI:  0.14 
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 : 3.83 
   Experiment series 25 Protein A Protein: DLG1  Fragment: 221-314  Site: PDZ_1  Construct: his6-MBP-TEVsite-PDZ 
Protein B Protein: RPS6KA1  Fragment: 725-735  Site: PBM  Modification: (TPO)734  Sequence: RVRKLPSTTL  Construct: Biotin-ttds-RRVRKLPSTpTL  
Average holdup BI:  0.11 
Normalized holdup BI:  0.14 
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 : 3.82 
   Experiment series 26 Protein A Protein: DLG1  Fragment: 457-570  Site: PDZ_3  Construct: his6-MBP-TEVsite-PDZ 
Protein B Protein: RPS6KA1  Fragment: 725-735  Site: PBM  Modification: (TPO)734  Sequence: RVRKLPSTTL  Construct: Biotin-ttds-RRVRKLPSTpTL  
Average holdup BI:  0.08 
Normalized holdup BI:  0.11 
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 : 3.7 
    
 
Please cite ProfAff! (Gogl, G. et al., Nature Communications 2022)  
© The ProfAff database is developped by the research team of Gergo Gogl .