Details of experiments between the fragments Q15418~725-735 and O95049~373-482
Note that the results of all experiments are listed, regardless of the modification states of the fragments.
Experiment series 1
Protein A protein: RPS6KA1
Protein A fragment: 725-735
Protein A site: PBM
Protein A modification: (SEP)732
Protein A sequence: RVRKLPSTTL
Protein A construct: biotin-ttds-RRVRKLPpSTTL
Protein B protein: TJP3
Protein B fragment: 373-482
Protein B site: PDZ_3
Protein B construct: his6-MBP-TEVsite-PDZ
Average holdup BI: 0.01
Holdup BI standard deviation: 0.15
Immobilized partner concentration in holdup experiment (10-6M): 40
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: 3
The affinity was below the detection threshold of the assay.
Experiment series 2
Protein A protein: RPS6KA1
Protein A fragment: 725-735
Protein A site: PBM
Protein A sequence: RVRKLPSTTL
Protein A construct: biotin-ttds-RRVRKLPSTTL
Protein B protein: TJP3
Protein B fragment: 373-482
Protein B site: PDZ_3
Protein B construct: his6-MBP-TEVsite-PDZ
Average holdup BI: 0.04
Holdup BI standard deviation: 0.01
Immobilized partner concentration in holdup experiment (10-6M): 31
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: 4
The affinity was below the detection threshold of the assay.
Experiment series 3
Protein A protein: RPS6KA1
Protein A fragment: 725-735
Protein A site: PBM
Protein A modification: (SEP)732
Protein A sequence: RVRKLPSTTL
Protein A construct: biotin-ttds-RRVRKLPpSTTL
Protein B protein: TJP3
Protein B fragment: 373-482
Protein B site: PDZ_3
Protein B construct: his6-MBP-TEVsite-PDZ
Average holdup BI: -0.07
Normalized holdup BI: -0.07
Immobilized partner concentration in holdup experiment (10-6M): 15
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 4
Protein A protein: RPS6KA1
Protein A fragment: 725-735
Protein A site: PBM
Protein A modification: (TPO)733
Protein A sequence: RVRKLPSTTL
Protein A construct: biotin-ttds-RRVRKLPSpTTL
Protein B protein: TJP3
Protein B fragment: 373-482
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): 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 5
Protein A protein: RPS6KA1
Protein A fragment: 725-735
Protein A site: PBM
Protein A modification: (TPO)734
Protein A sequence: RVRKLPSTTL
Protein A construct: Biotin-ttds-RRVRKLPSTpTL
Protein B protein: TJP3
Protein B fragment: 373-482
Protein B site: PDZ_3
Protein B construct: his6-MBP-TEVsite-PDZ
Average holdup BI: 0.03
Normalized holdup BI: 0.03
Immobilized partner concentration in holdup experiment (10-6M): 25
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 6
Protein A protein: RPS6KA1
Protein A fragment: 725-735
Protein A site: PBM
Protein A sequence: RVRKLPSTTL
Protein A construct: biotin-ttds-RRVRKLPSTTL
Protein B protein: TJP3
Protein B fragment: 373-482
Protein B site: PDZ_3
Protein B construct: his6-MBP-TEVsite-PDZ
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
PUBMED ID of publication: 36115835
Number of measurements: 1
Measured pKd: 3.59
Experiment series 7
Protein A protein: RPS6KA1
Protein A fragment: 725-735
Protein A site: PBM
Protein A modification: (SEP)732
Protein A sequence: RVRKLPSTTL
Protein A construct: biotin-ttds-RRVRKLPpSTTL
Protein B protein: TJP3
Protein B fragment: 373-482
Protein B site: PDZ_3
Protein B construct: his6-MBP-TEVsite-PDZ
Average holdup BI: 0.03
Normalized holdup BI: 0.03
Immobilized partner concentration in holdup experiment (10-6M): 15
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
The affinity was below the detection threshold of the assay.