Details of experiments between Q86UL8 and Q96DR7
Note that the results of all experiments are listed, regardless of the modification states of the fragments.
Experiment series 1
Protein A protein: MAGI2
Protein A fragment: 1141-1229
Protein A site: PDZ_6
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: ARHGEF26
Protein B fragment: 862-871
Protein B site: PBM
Protein B sequence: GRLLGLETNV
Protein B construct: biotin-ttds-GRLLGLETNV
Average holdup BI: 0.24
Holdup BI standard deviation: 0
Normalized holdup BI: 0.29
Normalized holdup BI standard deviation: 0
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: 2
Measured pKd: 4.39
Experiment series 2
Protein A protein: MAGI2
Protein A fragment: 772-865
Protein A site: PDZ_4
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: ARHGEF26
Protein B fragment: 862-871
Protein B site: PBM
Protein B sequence: GRLLGLETNV
Protein B construct: biotin-ttds-GRLLGLETNV
Average holdup BI: 0.11
Holdup BI standard deviation: 0
Normalized holdup BI: 0.15
Normalized holdup BI standard deviation: 0
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: 2
Measured pKd: 4.02
Experiment series 3
Protein A protein: MAGI2
Protein A fragment: 600-682
Protein A site: PDZ_3
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: ARHGEF26
Protein B fragment: 862-871
Protein B site: PBM
Protein B sequence: GRLLGLETNV
Protein B construct: biotin-ttds-GRLLGLETNV
Average holdup BI: 0.1
Holdup BI standard deviation: 0
Normalized holdup BI: 0.12
Normalized holdup BI standard deviation: 0
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: 2
Measured pKd: 3.9
Experiment series 4
Protein A protein: MAGI2
Protein A fragment: 410-522
Protein A site: PDZ_2
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: ARHGEF26
Protein B fragment: 862-871
Protein B site: PBM
Protein B sequence: GRLLGLETNV
Protein B construct: biotin-ttds-GRLLGLETNV
Average holdup BI: 0.08
Normalized holdup BI: 0.08
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: 3.72
Experiment series 5
Protein A protein: MAGI2
Protein A fragment: 915-1017
Protein A site: PDZ_5
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: ARHGEF26
Protein B fragment: 862-871
Protein B site: PBM
Protein B sequence: GRLLGLETNV
Protein B construct: biotin-ttds-GRLLGLETNV
Average holdup BI: 0.02
Holdup BI standard deviation: 0.01
Normalized holdup BI: 0.02
Normalized holdup BI standard deviation: 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: 2
The affinity was below the detection threshold of the assay.
Experiment series 6
Protein A protein: MAGI2
Protein A fragment: 410-522
Protein A site: PDZ_2
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: ARHGEF26
Protein B fragment: 862-871
Protein B site: PBM
Protein B sequence: GRLLGLETNV
Protein B construct: Biotin-ado-ado-GRLLGLETNV
Average holdup BI: 0.28
Normalized holdup BI: 0.32
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: Crude peptide synthesis product. The peptide concentration in the affinity conversion step may differ from reality. Not reported internal standards of holdup layout #6
PUBMED ID of publication: 36115835
Number of measurements: 1
Measured pKd: 4.44
Experiment series 7
Protein A protein: MAGI2
Protein A fragment: 1141-1229
Protein A site: PDZ_6
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: ARHGEF26
Protein B fragment: 862-871
Protein B site: PBM
Protein B sequence: GRLLGLETNV
Protein B construct: Biotin-ado-ado-GRLLGLETNV
Average holdup BI: 0.3
Normalized holdup BI: 0.3
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: Crude peptide synthesis product. The peptide concentration in the affinity conversion step may differ from reality. Not reported internal standards of holdup layout #6
PUBMED ID of publication: 36115835
Number of measurements: 1
Measured pKd: 4.4
Experiment series 8
Protein A protein: MAGI2
Protein A fragment: 600-682
Protein A site: PDZ_3
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: ARHGEF26
Protein B fragment: 862-871
Protein B site: PBM
Protein B sequence: GRLLGLETNV
Protein B construct: Biotin-ado-ado-GRLLGLETNV
Average holdup BI: 0.08
Normalized holdup BI: 0.08
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: Crude peptide synthesis product. The peptide concentration in the affinity conversion step may differ from reality. Not reported internal standards of holdup layout #6
PUBMED ID of publication: 36115835
Number of measurements: 1
Measured pKd: 3.72
Experiment series 9
Protein A protein: MAGI2
Protein A fragment: 772-865
Protein A site: PDZ_4
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: ARHGEF26
Protein B fragment: 862-871
Protein B site: PBM
Protein B sequence: GRLLGLETNV
Protein B construct: Biotin-ado-ado-GRLLGLETNV
Average holdup BI: 0.11
Normalized holdup BI: 0.14
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: Crude peptide synthesis product. The peptide concentration in the affinity conversion step may differ from reality. Not reported internal standards of holdup layout #6
PUBMED ID of publication: 36115835
Number of measurements: 1
Measured pKd: 3.98
Experiment series 10
Protein A protein: MAGI2
Protein A fragment: 915-1017
Protein A site: PDZ_5
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: ARHGEF26
Protein B fragment: 862-871
Protein B site: PBM
Protein B sequence: GRLLGLETNV
Protein B construct: Biotin-ado-ado-GRLLGLETNV
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
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
PUBMED ID of publication: 36115835
Number of measurements: 1
The affinity was below the detection threshold of the assay.