Details of experiments between Q96HA1 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: POM121
Protein A fragment: 1-1249
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: 0.21
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): 0.97
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: POM121
Protein A fragment: 140-154
Protein A site: PxxP_1
Protein A sequence: YLGKPGPPQPAAAPE
Protein A construct: Bio Ado Ado-YLGKPGPPQPAAAPE-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
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 3

Protein A protein: POM121
Protein A fragment: 161-175
Protein A site: PxxP_2
Protein A sequence: RPGRRPPARPAPRSP
Protein A construct: Bio Ado Ado-RPGRRPPARPAPRSP-CONH2

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

Average holdup BI: -0.01
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: 1
The affinity was below the detection threshold of the assay.


Experiment series 4

Protein A protein: POM121
Protein A fragment: 166-180
Protein A site: PxxP_3
Protein A sequence: PPARPAPRSPPPRSP
Protein A construct: Bio Ado Ado-PPARPAPRSPPPRSP-CONH2

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

Average holdup BI: -0.01
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: 1
The affinity was below the detection threshold of the assay.


Experiment series 5

Protein A protein: POM121
Protein A fragment: 180-194
Protein A site: PxxP_4
Protein A sequence: PPPRSPPPSPPTHRA
Protein A construct: Bio Ado Ado-PPPRSPPPSPPTHRA-CONH2

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

Average holdup BI: -0.05
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 6

Protein A protein: POM121
Protein A fragment: 442-456
Protein A site: PxxP_5
Protein A sequence: SSRSQTPERPAKKIR
Protein A construct: Bio Ado Ado-SSRSQTPERPAKKIR-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.


Experiment series 7

Protein A protein: POM121
Protein A fragment: 737-751
Protein A site: PxxP_6
Protein A sequence: KSEKEGPTPPGPSVT
Protein A construct: Bio Ado Ado-KSEKEGPTPPGPSVT-CONH2

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

Average holdup BI: -0.04
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.