Details of experiments between P06400 and P00519
Note that the results of all experiments are listed, regardless of the modification states of the fragments.
Experiment series 1
Protein A protein: RB1
Protein A fragment: 1-928
Protein A construct: endogenous protein (Jurkat)
Protein B protein: ABL1
Protein B fragment: 56-121
Protein B site: SH3
Protein B construct: avi(biotin)-his6-MBP-TEVsite-SH3
Average holdup BI: 0.02
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): 1.45
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: RB1
Protein A fragment: 244-258
Protein A site: PxxP_1
Protein A sequence: IPINGSPRTPRRGQN
Protein A construct: Biotinylated 15-mer peptide (crude)
Protein B protein: ABL1
Protein B fragment: 56-121
Protein B site: SH3
Protein B construct: avi(biotin)-his6-MBP-TEVsite-SH3
Average holdup BI: -0
Holdup BI standard deviation: 0.01
Immobilized partner concentration in holdup experiment (10-6M): 18
Experiment method: HU (DAPF)
Date of measurement: 2022.06.29.
Number of measurements: 2
The affinity was below the detection threshold of the assay.
Experiment series 3
Protein A protein: RB1
Protein A fragment: 365-379
Protein A site: PxxP_2
Protein A sequence: EVNVIPPHTPVRTVM
Protein A construct: Biotinylated 15-mer peptide (crude)
Protein B protein: ABL1
Protein B fragment: 56-121
Protein B site: SH3
Protein B construct: avi(biotin)-his6-MBP-TEVsite-SH3
Average holdup BI: 0.01
Holdup BI standard deviation: 0.03
Immobilized partner concentration in holdup experiment (10-6M): 18
Experiment method: HU (DAPF)
Date of measurement: 2022.06.29.
Number of measurements: 2
The affinity was below the detection threshold of the assay.
Experiment series 4
Protein A protein: RB1
Protein A fragment: 780-794
Protein A site: PxxP_3
Protein A sequence: SPIPHIPRSPYKFPS
Protein A construct: Biotinylated 15-mer peptide (crude)
Protein B protein: ABL1
Protein B fragment: 56-121
Protein B site: SH3
Protein B construct: avi(biotin)-his6-MBP-TEVsite-SH3
Average holdup BI: 0.06
Holdup BI standard deviation: 0
Immobilized partner concentration in holdup experiment (10-6M): 18
Experiment method: HU (DAPF)
Date of measurement: 2022.06.29.
Number of measurements: 2
Measured pKd: 3.59
Experiment series 5
Protein A protein: RB1
Protein A fragment: 787-801
Protein A site: PxxP_4
Protein A sequence: RSPYKFPSSPLRIPG
Protein A construct: Biotinylated 15-mer peptide (crude)
Protein B protein: ABL1
Protein B fragment: 56-121
Protein B site: SH3
Protein B construct: avi(biotin)-his6-MBP-TEVsite-SH3
Average holdup BI: 0.05
Holdup BI standard deviation: 0.03
Immobilized partner concentration in holdup experiment (10-6M): 18
Experiment method: HU (DAPF)
Date of measurement: 2022.06.29.
Number of measurements: 2
Measured pKd: 3.5
Experiment series 6
Protein A protein: RB1
Protein A fragment: 862-876
Protein A site: PxxP_5
Protein A sequence: SAEGSNPPKPLKKLR
Protein A construct: Biotinylated 15-mer peptide (crude)
Protein B protein: ABL1
Protein B fragment: 56-121
Protein B site: SH3
Protein B construct: avi(biotin)-his6-MBP-TEVsite-SH3
Average holdup BI: -0
Holdup BI standard deviation: 0
Immobilized partner concentration in holdup experiment (10-6M): 18
Experiment method: HU (DAPF)
Date of measurement: 2022.06.29.
Number of measurements: 2
The affinity was below the detection threshold of the assay.