Details of experiments between Q8TBC3 and Q86UL8
Note that the results of all experiments are listed, regardless of the modification states of the fragments.
Experiment series 1
Protein A protein: SHKBP1
Protein A fragment: 698-707
Protein A site: PBM
Protein A sequence: PKMKLNETSF
Protein A construct: Biotin-Ahx-PKMKLNETSF
Protein B protein: MAGI2
Protein B fragment: 772-865
Protein B site: PDZ_4
Protein B construct: his6-MBP-TEVsite-PDZ
Average holdup BI: 0.07
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: The peptide stock solution had solubility issues. The peptide concentration in the affinity conversion step may differ from reality causing a systematic error in reported affinities.
PUBMED ID of publication: 36115835
Number of measurements: 1
Measured pKd: 3.69
Experiment series 2
Protein A protein: SHKBP1
Protein A fragment: 698-707
Protein A site: PBM
Protein A sequence: PKMKLNETSF
Protein A construct: Biotin-Ahx-PKMKLNETSF
Protein B protein: MAGI2
Protein B fragment: 1141-1229
Protein B site: PDZ_6
Protein B construct: his6-MBP-TEVsite-PDZ
Average holdup BI: 0.05
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: The peptide stock solution had solubility issues. The peptide concentration in the affinity conversion step may differ from reality causing a systematic error in reported affinities.
PUBMED ID of publication: 36115835
Number of measurements: 1
Measured pKd: 3.54
Experiment series 3
Protein A protein: SHKBP1
Protein A fragment: 698-707
Protein A site: PBM
Protein A sequence: PKMKLNETSF
Protein A construct: Biotin-Ahx-PKMKLNETSF
Protein B protein: MAGI2
Protein B fragment: 915-1017
Protein B site: PDZ_5
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
Experimental details: The peptide stock solution had solubility issues. The peptide concentration in the affinity conversion step may differ from reality causing a systematic error in reported affinities.
PUBMED ID of publication: 36115835
Number of measurements: 1
Measured pKd: 3.53
Experiment series 4
Protein A protein: SHKBP1
Protein A fragment: 698-707
Protein A site: PBM
Protein A sequence: PKMKLNETSF
Protein A construct: Biotin-Ahx-PKMKLNETSF
Protein B protein: MAGI2
Protein B fragment: 410-522
Protein B site: PDZ_2
Protein B construct: his6-MBP-TEVsite-PDZ
Average holdup BI: -0.03
Holdup BI standard deviation: 0
Normalized holdup BI: -0.04
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
Experimental details: The peptide stock solution had solubility issues. The peptide concentration in the affinity conversion step may differ from reality causing a systematic error in reported affinities.
PUBMED ID of publication: 36115835
Number of measurements: 2
The affinity was below the detection threshold of the assay.
Experiment series 5
Protein A protein: SHKBP1
Protein A fragment: 698-707
Protein A site: PBM
Protein A sequence: PKMKLNETSF
Protein A construct: Biotin-Ahx-PKMKLNETSF
Protein B protein: MAGI2
Protein B fragment: 600-682
Protein B site: PDZ_3
Protein B construct: his6-MBP-TEVsite-PDZ
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
Experimental details: The peptide stock solution had solubility issues. The peptide concentration in the affinity conversion step may differ from reality causing a systematic error in reported affinities.
PUBMED ID of publication: 36115835
Number of measurements: 1
The affinity was below the detection threshold of the assay.