Details of experiments between P49418 and P46013
Note that the results of all experiments are listed, regardless of the modification states of the fragments.
Experiment series 1
Protein A protein: AMPH
Protein A fragment: 615-695
Protein A site: SH3
Protein A construct: avi(biotin)-his6-MBP-TEVsite-SH3
Protein B protein: MKI67
Protein B fragment: 1-3256
Protein B construct: endogenous protein (Jurkat)
Average holdup BI: 0.34
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): 6.47
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: 4.71
Experiment series 2
Protein A protein: AMPH
Protein A fragment: 615-695
Protein A site: SH3
Protein A construct: avi(biotin)-his6-MBP-TEVsite-SH3
Protein B protein: MKI67
Protein B fragment: 1083-1097
Protein B site: PxxP_1
Protein B sequence: TGMKKWPRTPKEEAQ
Protein B construct: Biotinylated 15-mer peptide (crude)
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: AMPH
Protein A fragment: 615-695
Protein A site: SH3
Protein A construct: avi(biotin)-his6-MBP-TEVsite-SH3
Protein B protein: MKI67
Protein B fragment: 1327-1341
Protein B site: PxxP_2
Protein B sequence: TGSKRRPQTPKEEAQ
Protein B construct: Biotinylated 15-mer peptide (crude)
Average holdup BI: -0.03
Holdup BI standard deviation: 0.02
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: AMPH
Protein A fragment: 615-695
Protein A site: SH3
Protein A construct: avi(biotin)-his6-MBP-TEVsite-SH3
Protein B protein: MKI67
Protein B fragment: 1385-1399
Protein B site: PxxP_3
Protein B sequence: TSTRRQPKTPLEKRD
Protein B construct: Biotinylated 15-mer peptide (crude)
Average holdup BI: 0.04
Holdup BI standard deviation: 0.02
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 5
Protein A protein: AMPH
Protein A fragment: 615-695
Protein A site: SH3
Protein A construct: avi(biotin)-his6-MBP-TEVsite-SH3
Protein B protein: MKI67
Protein B fragment: 1935-1949
Protein B site: PxxP_4
Protein B sequence: PGSKRRPQTPKEKAK
Protein B construct: Biotinylated 15-mer peptide (crude)
Average holdup BI: 0.03
Holdup BI standard deviation: 0.02
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 6
Protein A protein: AMPH
Protein A fragment: 615-695
Protein A site: SH3
Protein A construct: avi(biotin)-his6-MBP-TEVsite-SH3
Protein B protein: MKI67
Protein B fragment: 2115-2129
Protein B site: PxxP_5
Protein B sequence: TSTRRRPKTPLGKRD
Protein B construct: Biotinylated 15-mer peptide (crude)
Average holdup BI: 0.11
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: 3.86
Experiment series 7
Protein A protein: AMPH
Protein A fragment: 615-695
Protein A site: SH3
Protein A construct: avi(biotin)-his6-MBP-TEVsite-SH3
Protein B protein: MKI67
Protein B fragment: 2175-2189
Protein B site: PxxP_6
Protein B sequence: TGSKRQPRTPKGKAQ
Protein B construct: Biotinylated 15-mer peptide (crude)
Average holdup BI: 0.03
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.
Experiment series 8
Protein A protein: AMPH
Protein A fragment: 615-695
Protein A site: SH3
Protein A construct: avi(biotin)-his6-MBP-TEVsite-SH3
Protein B protein: MKI67
Protein B fragment: 2297-2311
Protein B site: PxxP_7
Protein B sequence: PGSKRWPQTPKEKAQ
Protein B construct: Biotinylated 15-mer peptide (crude)
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 9
Protein A protein: AMPH
Protein A fragment: 615-695
Protein A site: SH3
Protein A construct: avi(biotin)-his6-MBP-TEVsite-SH3
Protein B protein: MKI67
Protein B fragment: 2418-2432
Protein B site: PxxP_8
Protein B sequence: PGSKRQPQTPKEKAE
Protein B construct: Biotinylated 15-mer peptide (crude)
Average holdup BI: -0.03
Holdup BI standard deviation: 0.02
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 10
Protein A protein: AMPH
Protein A fragment: 615-695
Protein A site: SH3
Protein A construct: avi(biotin)-his6-MBP-TEVsite-SH3
Protein B protein: MKI67
Protein B fragment: 2660-2674
Protein B site: PxxP_9
Protein B sequence: EPSRRRPRAPKEKAQ
Protein B construct: Biotinylated 15-mer peptide (crude)
Average holdup BI: 0.08
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
Measured pKd: 3.66
Experiment series 11
Protein A protein: AMPH
Protein A fragment: 615-695
Protein A site: SH3
Protein A construct: avi(biotin)-his6-MBP-TEVsite-SH3
Protein B protein: MKI67
Protein B fragment: 2781-2795
Protein B site: PxxP_10
Protein B sequence: TGSRRRPRAPRESAQ
Protein B construct: Biotinylated 15-mer peptide (crude)
Average holdup BI: 0.12
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: 3.89
Experiment series 12
Protein A protein: AMPH
Protein A fragment: 615-695
Protein A site: SH3
Protein A construct: avi(biotin)-his6-MBP-TEVsite-SH3
Protein B protein: MKI67
Protein B fragment: 2899-2913
Protein B site: PxxP_11
Protein B sequence: IGSRRQPRAPKEKAQ
Protein B construct: Biotinylated 15-mer peptide (crude)
Average holdup BI: 0.03
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 13
Protein A protein: AMPH
Protein A fragment: 615-695
Protein A site: SH3
Protein A construct: avi(biotin)-his6-MBP-TEVsite-SH3
Protein B protein: MKI67
Protein B fragment: 2986-3000
Protein B site: PxxP_12
Protein B sequence: KSNTSLPPLPFKRGG
Protein B construct: Biotinylated 15-mer peptide (crude)
Average holdup BI: 0.05
Holdup BI standard deviation: 0.05
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 14
Protein A protein: AMPH
Protein A fragment: 615-695
Protein A site: SH3
Protein A construct: avi(biotin)-his6-MBP-TEVsite-SH3
Protein B protein: MKI67
Protein B fragment: 517-531
Protein B site: PxxP_13
Protein B sequence: FDENLPPNTPLKRGE
Protein B construct: Biotinylated 15-mer peptide (crude)
Average holdup BI: -0.06
Holdup BI standard deviation: 0.05
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 15
Protein A protein: AMPH
Protein A fragment: 615-695
Protein A site: SH3
Protein A construct: avi(biotin)-his6-MBP-TEVsite-SH3
Protein B protein: MKI67
Protein B fragment: 686-700
Protein B site: PxxP_14
Protein B sequence: NKRQRRPATPKKPVG
Protein B construct: Biotinylated 15-mer peptide (crude)
Average holdup BI: 0.07
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: 3.62