Details of experiments between P49418 and Q8IYB3
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: SRRM1
Protein B fragment: 1-904
Protein B construct: endogenous protein (Jurkat)
Average holdup BI: 0.1
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): 4.46
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: AMPH
Protein A fragment: 615-695
Protein A site: SH3
Protein A construct: avi(biotin)-his6-MBP-TEVsite-SH3
Protein B protein: SRRM1
Protein B fragment: 327-341
Protein B site: PxxP_1
Protein B sequence: TPPRRMPPPPRHRRS
Protein B construct: Biotinylated 15-mer peptide (crude)
Average holdup BI: 0.47
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
Measured pKd: 4.75
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: SRRM1
Protein B fragment: 365-379
Protein B site: PxxP_2
Protein B sequence: SRSRSPPKKPPKRTS
Protein B construct: Biotinylated 15-mer peptide (crude)
Average holdup BI: 0.24
Holdup BI standard deviation: 0.04
Immobilized partner concentration in holdup experiment (10-6M): 18
Experiment method: HU (DAPF)
Date of measurement: 2022.06.29.
Number of measurements: 4
Measured pKd: 4.27
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: SRRM1
Protein B fragment: 395-409
Protein B site: PxxP_3
Protein B sequence: PRRRHRPSPPATPPP
Protein B construct: Biotinylated 15-mer peptide (crude)
Average holdup BI: 0.3
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
Measured pKd: 4.41
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: SRRM1
Protein B fragment: 544-558
Protein B site: PxxP_4
Protein B sequence: GRRRRSPSPPPTRRR
Protein B construct: Biotinylated 15-mer peptide (crude)
Average holdup BI: 0.67
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: 5.13
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: SRRM1
Protein B fragment: 557-571
Protein B site: PxxP_5
Protein B sequence: RRRSPSPAPPPRRRR
Protein B construct: Biotinylated 15-mer peptide (crude)
Average holdup BI: 0.84
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: 5.55
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: SRRM1
Protein B fragment: 567-581
Protein B site: PxxP_6
Protein B sequence: PRRRRTPTPPPRRRT
Protein B construct: Biotinylated 15-mer peptide (crude)
Average holdup BI: 0.86
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: 4
Measured pKd: 5.63
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: SRRM1
Protein B fragment: 576-590
Protein B site: PxxP_7
Protein B sequence: PPRRRTPSPPPRRRS
Protein B construct: Biotinylated 15-mer peptide (crude)
Average holdup BI: 0.81
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: 4
Measured pKd: 5.46
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: SRRM1
Protein B fragment: 600-614
Protein B site: PxxP_8
Protein B sequence: IQRRYSPSPPPKRRT
Protein B construct: Biotinylated 15-mer peptide (crude)
Average holdup BI: 0.3
Holdup BI standard deviation: 0.08
Immobilized partner concentration in holdup experiment (10-6M): 18
Experiment method: HU (DAPF)
Date of measurement: 2022.06.29.
Number of measurements: 4
Measured pKd: 4.41
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: SRRM1
Protein B fragment: 611-625
Protein B site: PxxP_9
Protein B sequence: KRRTASPPPPPKRRA
Protein B construct: Biotinylated 15-mer peptide (crude)
Average holdup BI: 0.32
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: 4.44
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: SRRM1
Protein B fragment: 612-626
Protein B site: PxxP_10
Protein B sequence: RRTASPPPPPKRRAS
Protein B construct: Biotinylated 15-mer peptide (crude)
Average holdup BI: 0.36
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
Measured pKd: 4.53
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: SRRM1
Protein B fragment: 621-635
Protein B site: PxxP_11
Protein B sequence: PKRRASPSPPPKRRV
Protein B construct: Biotinylated 15-mer peptide (crude)
Average holdup BI: 0.38
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: 4
Measured pKd: 4.56
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: SRRM1
Protein B fragment: 759-773
Protein B site: PxxP_12
Protein B sequence: PAAKKPPAPPSPVQS
Protein B construct: Biotinylated 15-mer peptide (crude)
Average holdup BI: 0.01
Holdup BI standard deviation: 0.04
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 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: SRRM1
Protein B fragment: 776-790
Protein B site: PxxP_13
Protein B sequence: PSTNWSPAVPVKKAK
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 15
Protein A protein: AMPH
Protein A fragment: 615-695
Protein A site: SH3
Protein A construct: his6-MBP-TEVsite-SH3
Protein B protein: SRRM1
Protein B fragment: 576-590
Protein B site: PxxP_7
Protein B sequence: PPRRRTPSPPPRRRS
Protein B construct: Bio E E Ado Ado-PPRRRTPSPPPRRRS-CONH2
Average holdup BI: 0.32
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: 4.46