Details of experiments between Q99501 and P49418
Note that the results of all experiments are listed, regardless of the modification states of the fragments.
Experiment series 1
Protein A protein: GAS2L1
Protein A fragment: 1-681
Protein A construct: endogenous protein (Jurkat)
Protein B protein: AMPH
Protein B fragment: 615-695
Protein B site: SH3
Protein B construct: avi(biotin)-his6-MBP-TEVsite-SH3
Average holdup BI: 0.73
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): 3.11
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
Measured pKd: 5.43
Experiment series 2
Protein A protein: GAS2L1
Protein A fragment: 166-180
Protein A site: PxxP_1
Protein A sequence: RELRAAPPAPNAPAA
Protein A construct: Bio Ado Ado-RELRAAPPAPNAPAA-CONH2
Protein B protein: AMPH
Protein B fragment: 615-695
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: 2023/01/16
Number of measurements: 1
Measured pKd: 3.49
Experiment series 3
Protein A protein: GAS2L1
Protein A fragment: 185-199
Protein A site: PxxP_2
Protein A sequence: TETAPAPGTPARGPR
Protein A construct: Bio Ado Ado-TETAPAPGTPARGPR-CONH2
Protein B protein: AMPH
Protein B fragment: 615-695
Protein B site: SH3
Protein B construct: his6-MBP-TEVsite-SH3
Average holdup BI: 0.1
Immobilized partner concentration in holdup experiment (10-6M): 18
Experiment method: HU (DAPF)
Details of affinity fitting: Quadratic binding equation
Date of measurement: 2023/01/16
Number of measurements: 1
Measured pKd: 3.78
Experiment series 4
Protein A protein: GAS2L1
Protein A fragment: 326-340
Protein A site: PxxP_3
Protein A sequence: RSTKEGPETPPRPRD
Protein A construct: Bio Ado Ado-RSTKEGPETPPRPRD-CONH2
Protein B protein: AMPH
Protein B fragment: 615-695
Protein B site: SH3
Protein B construct: his6-MBP-TEVsite-SH3
Average holdup BI: 0.02
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: 2023/01/16
Number of measurements: 2
The affinity was below the detection threshold of the assay.
Experiment series 5
Protein A protein: GAS2L1
Protein A fragment: 386-400
Protein A site: PxxP_4
Protein A sequence: RLTTGTPASPRRPPA
Protein A construct: Bio Ado Ado-RLTTGTPASPRRPPA-CONH2
Protein B protein: AMPH
Protein B fragment: 615-695
Protein B site: SH3
Protein B construct: his6-MBP-TEVsite-SH3
Average holdup BI: -0.09
Immobilized partner concentration in holdup experiment (10-6M): 18
Experiment method: HU (DAPF)
Details of affinity fitting: Quadratic binding equation
Date of measurement: 2023/01/16
Number of measurements: 1
The affinity was below the detection threshold of the assay.
Experiment series 6
Protein A protein: GAS2L1
Protein A fragment: 466-480
Protein A site: PxxP_5
Protein A sequence: GSGRSTPQTPRARSP
Protein A construct: Bio Ado Ado-GSGRSTPQTPRARSP-CONH2
Protein B protein: AMPH
Protein B fragment: 615-695
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: 2023/01/16
Number of measurements: 1
The affinity was below the detection threshold of the assay.
Experiment series 7
Protein A protein: GAS2L1
Protein A fragment: 644-658
Protein A site: PxxP_6
Protein A sequence: PSRIPTPRGPRRPSG
Protein A construct: Bio Ado Ado-PSRIPTPRGPRRPSG-CONH2
Protein B protein: AMPH
Protein B fragment: 615-695
Protein B site: SH3
Protein B construct: his6-MBP-TEVsite-SH3
Average holdup BI: 0.06
Immobilized partner concentration in holdup experiment (10-6M): 18
Experiment method: HU (DAPF)
Details of affinity fitting: Quadratic binding equation
Date of measurement: 2023/01/16
Number of measurements: 1
Measured pKd: 3.57