Details of experiments between Q96DX4 and Q5VST9
Note that the results of all experiments are listed, regardless of the modification states of the fragments.



Experiment series 1

Protein A protein: RSPRY1
Protein A fragment: 1-576
Protein A construct: endogenous protein (Jurkat)

Protein B protein: OBSCN
Protein B fragment: 5594-5674
Protein B site: SH3
Protein B construct: avi(biotin)-his6-MBP-TEVsite-SH3

Average holdup BI: -0.11
Immobilized partner concentration in holdup experiment (10-6M): 10
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 0.41
Experiment method: nHU_Jurkat
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022.06.29.
Experimental details: -log10(P Two-sided unpaired T-test (6 vs 3))
Number of measurements: 3
The affinity was below the detection threshold of the assay.


Experiment series 2

Protein A protein: RSPRY1
Protein A fragment: 71-85
Protein A site: PxxP_1
Protein A sequence: ADTRSQPRDPVRPPR
Protein A construct: Bio Ado Ado-ADTRSQPRDPVRPPR-CONH2

Protein B protein: OBSCN
Protein B fragment: 5594-5674
Protein B site: SH3
Protein B construct: his6-MBP-TEVsite-SH3

Average holdup BI: -0.03
Holdup BI standard deviation: 0.01
Immobilized partner concentration in holdup experiment (10-6M): 18
Experiment method: HU (DAPF)
Details of affinity fitting: Quadratic binding equation
Date of measurement: 2023/01/18
Number of measurements: 2
The affinity was below the detection threshold of the assay.


Experiment series 3

Protein A protein: RSPRY1
Protein A fragment: 74-88
Protein A site: PxxP_2
Protein A sequence: RSQPRDPVRPPRRGR
Protein A construct: Bio Ado Ado-RSQPRDPVRPPRRGR-CONH2

Protein B protein: OBSCN
Protein B fragment: 5594-5674
Protein B site: SH3
Protein B construct: his6-MBP-TEVsite-SH3

Average holdup BI: -0.08
Immobilized partner concentration in holdup experiment (10-6M): 18
Experiment method: HU (DAPF)
Details of affinity fitting: Quadratic binding equation
Date of measurement: 2023/01/18
Number of measurements: 1
The affinity was below the detection threshold of the assay.


Experiment series 4

Protein A protein: RSPRY1
Protein A fragment: 84-98
Protein A site: PxxP_3
Protein A sequence: PRRGRGPHEPRRKKQ
Protein A construct: Bio Ado Ado-PRRGRGPHEPRRKKQ-CONH2

Protein B protein: OBSCN
Protein B fragment: 5594-5674
Protein B site: SH3
Protein B construct: his6-MBP-TEVsite-SH3

Average holdup BI: -0.02
Immobilized partner concentration in holdup experiment (10-6M): 18
Experiment method: HU (DAPF)
Details of affinity fitting: Quadratic binding equation
Date of measurement: 2023/01/18
Number of measurements: 1
The affinity was below the detection threshold of the assay.