Dear MST User,
In the following you find a list of interactions/molecule classes that have been analyzed with MST as well as with other methods. Thus, this compilation does
not reflect the whole spectrum of systems that have been measured with MST so far. The list gives detailed information on instrument settings and assay
conditions to also support your assay development.
Antibody - Antigen
Target Name
Ligand
MST Kd
Orthogonal Kd
Litera- Buffer
ture
MST Conditions
mAB08
mAB08
Cocaine, no serum
Cocaine, 20 %
serum
Benzoylegonine
Cocaethylene
BSA-SNC
7.8 nM
105 nM
2.5 ± 6.8 nM (ITC)
113 ± 820 nM (ITC)
(1)
PBS + 0.05 % Tween-20
NT.115
standard treated capillaries
NT-647
23.7 nM
47 nM
6.6 nM
18.6 ± 48.7 nM (ITC)
34.4 ± 129 nM (ITC)
-
MB007-antibody
10 ± 0.6 nM
14.9 ± 0.8 nM
(IC50 in Cell Assay)
(2)
50 mM sodium phosphate pH 7.4,
150 mM NaCl, 0.05 % Tween-20
NT.115
standard treated capillaries
30 % LED, 25 % MST
mAB08
mAB08
mAB08
GM-CSF
Cytokine
1
Protein - protein
Target Name
Ligand
MST Kd
Orthogonal Kd
Litera- Buffer
ture
MST Conditions
Β-Lactamase
TEM1
WT BLIP
3.5 ± 0.6 nM
(3,4)
W112A BLIP
W150A BLIP
474 ± 76 nM
1750 ± 220 nM
3.2 ± 0.6 nM
(SPR)
360 ± 60 nM
3800 ± 600 nM
NT.115
standard treated capillaries
50 % LED, 80 % MST
Grb2
Grb2
0.66 ± 0.20 µM
µM range (DLS)
(5)
NEMO UBAN-ZF
NEMO UBAN-ZF
NEMO UBAN-ZF
linear diUbi
K63-linked diUbi
linear tetraUbi
3.24 µM
No binding
0.34 µM
1.4 µM (ITC)
131 µM (ITC)
Western Blot –
not quantifiable
(6,7)
(6,7)
(7)
50 mM Tris-HCl, pH 7.6, 150 mM
NaCl, 10 mM MgCl2, 0.05 % Tween-20
NT.115
10 % LED, 40 % MST
NT-647
25 mM Tris-HCl pH 8.0, 100 mM NaCl,
0.1 % BSA, 0.1 % Tween-20, 0.5 mM
DTT
NT.115
NT-647
Protein - Ion
Target Name
Ligand
MST Kd
Orthogonal Kd
Literature
Buffer
MST Conditions
Calmodulin
M13
4.48 ± 1.7 nM
1.6 nM
(22)
PBS +0.025 % Tween-20
Calcium
2.8 ± 0.2 µM
µM-range
(23)
1 × PBS
NT.115
standard treated capillaries
80 % LED, 20 % MST
NT.115
2
Protein -Small molecule
Target Name
Ligand
MST Kd
Orthogonal Kd
Literature
Buffer
MST Conditions
Hsp90
17-DMAG
0.503 ±
0.099 μM
0.35 ± 0.04 μM
(ITC)
(8,9)
50 mM Tris-HCl, pH 7.4, 150 mM
NaCl, 10 mM MgCl2, 0.05 % (v/v)
Tween-20, 5 % (v/v) ethanol
NT.115
hydrophilic treated capillaries
40 % LED, 40 % MST
NT-647
NT.LabelFree
standard treated capillaries
20 % MST
NT.115
standard treated capillaries
80 % LED, 40% MST
NT-647
NT.115
hydrophilic treated capillaries
80 % LED, 40 % MST
NT.115
standard treated capillaries
0.593 ±
0.387 μM
P38a
Kinase
P53
Kinase
BIRB796
2.3 ± 0.17 nM
0.1 nM
(10)
(stopped flow –
fluorescence)
2.5 % DMSO in MST-Buffer
SB203580
24 ± 4.37 nM
15 nM (ITC)
(11)
MST Buffer
PD169316
18 ± 15 nM
130 nM (IC50)
(SPR)
(12,13)
SB202190
SB239063
48 ± 21 nM
8 ± 12 nM
37 nM
44 nM (IC50)
(13,14)
(13,15)
50 mM Tris, 150 mM NaCl,
10 mM MgCl2, 0.05 % Tween-20,
pH 7.8
200 mM TAPS/Tris pH 8.0, 1 mM
DTT, 0.05 % Tween-20
Nutlin3
36.3 nM
FP-Based
Binding Assay
36 nM
(16)
79 % LED
20 % MST
3
Target Name
Ligand
MST Kd
Orthogonal Kd
Literature
MDM2
Kinase
P53
260 nM
50 - 300 nM
(17)
RhoA
GTPase
G04 (compound)
400 nM
Multiple Cell
based assays
(18)
Rac
GTPase
P67
31.6 nM
Activity
(19)
P67
Phox-I1
97.5 nM
Activity
(19)
Buffer
MST Conditions
50 mM HEPES, 50 mM NaCl, 0.01 %
Tween-20 and 2 mM MgCl2
NT.115
NT-647
NT.115
NT-647
4
Protein - Sugar
Target Name
Ligand
MST Kd
Orthogonal Kd
Literature
Buffer
MST Conditions
Nod factor
NFR5
4.0 ± 1.5 nM
(35)
0.6 ± 0.25 nM
50 mM Tris, pH 8, 0.2 M NaCl,
2 mM β-ME, 0.36 % (w/v) FosCholine
NT.115
100 % LED, 50 % MST
NFR1
10.1 ± 2.5 nM
(SPR)
4.9 ± 1.3 nM (SPR)
Riproximin
ASF
2 binding sites:
11 nM and 7 µM
2 binding sites:
48 nM and 1.1 µM
(ITC)
(36)
PBS with 50 mM galactose and
0.05 % Tween-20
NT.115
standard treated capillaries
NT-647
Lysozyme
Glc-NAc
18.8 ± 1.9 mM
25 mM (ITC)
(37)
MST-Buffer with GlcNAc
(dissolved in NaActetate Buffer)
NT.115
40 % MST
NT-488
FedF
Blood group A type
1 hexa- saccharide
NFR1
2.9 µM ± 1.6 µM
SPR (35.2 µM)
BSI (1.76 µM)
0.6 ± 0.25 nM
(38)
20 mM HEPES pH 7.4, 150 mM
NaCl, 0.005 % Tween-20
NT.115
NT-647
0.6 ± 0.25 nM
5
Protein - nucleic acids
Target Name
Ligand
MST Kd
Orthogonal Kd
Literature
Buffer
MST Conditions
Thrombinaptamer
Thrombin
30 ± 19 nM
25 ± 25 nM
(24,25)
20 mM Tris-HCl pH 7.4, 150 mM
NaCl, 5 mM KCl, 1 mM CaCl2, 1 mM
MgCl2, 0.01 % Tween-20, 4 % BSA
NT.115
NT-647
Thrombin
Fibrinogen aptamer
32 ± 15 nM
25 ± 25 nM
(24,26)
Heparin aptamer
133 ± 42 nM
0.5 – 237 nM
(SPR, SELEX,
ACE)
(26-29)
20 mM Tris-HCl pH 7.4,
NT.115
150 mM NaCl, 5 mM KCl, 1 mM
standard treated capillaries
CaCl2, 1 mM MgCl2, 0.1 % Tween-20
Pur-α
Transcription
Factor
CAG-Triplet
opa-Repeat
834 ± 179 nM
1406 ± 128 nM
716 ± 202 nM
1211 ± 128 nM
(Anisotropy)
(30)
DNA gyrase
4,5’-bithiazole
compounds
Cmp1: 51.5 µM
47.4 ± 8.8 µM
(SPR)
6.6 ± 2 µM
(SPR)
(31)
n/a
n/a
n/a
n/a
(32)
Cmp2: 7.9 µM
STAT3-EGFP
STAT3-EGFP
STAT3-EGFP
STAT3-EGFP
GAS
S+100
S+100 mutant1
S+100 mutant2
37.9 ± 1.0 µM
23.3 ± 0.6 µM
740 ± 21 µM
No binding
20mM Tris pH 7.4, 100 mM KAc,
3.5 mM MgCl2, 0.1 % BSA, 0.01 %
Tween-20
NT.115
NT.115
standard treated capillaries
90 % LED, 80 % MST
NT-647
25 mM HEPES, pH 7.2; 50 mM
NaCl; 2.5 mM MgCl2; 0.025 % NP40
NT.115
standard treated capillaries
50 % LED
6
Nucleic acids
Target Name
Ligand
MST Kd
Orthogonal Kd
Literature
Buffer
MST Conditions
ATP/AMPaptamer
ATP/AMP
60 ± 4 µM (ATP)
Coincidence
with literature
Fluorescence
spectroscopy
(33,34)
20 mM Tris-HCl pH 7.6, 300 mM
NaCl, 5 mM MgCl2, 0.01%
Tween-20
NT.115
NT-647
87 ± 5 µM
(AMP)
(25)
Membrane Proteins
Target Name
Ligand
MST Kd
Orthogonal Kd
Literature
Buffer
MST Conditions
Receptor GluA2LBD
Glutamate
835 ± 43 nM
821 nM
(fluorescence
spectroscopy)
(13,39)
10 mM HEPES pH 8.0, 150 NaCl,
1 mM EDTA
NT.LabelFree
standard treated capillaries
4-Glu-Azu
22 ± 8 µM
19 ± 5 µM
Calcium
221 ± 23 µM
Synaptotagmin-1
206 ± 40 µM
NT.115
NT.LabelFree
5 µM - 1 mM (ITC)
(40-42)
20 mM HEPES, 150 mM KCl,
2.5 mg/ml BSA, pH 7.4
20 mM HEPES, 150 mM KCl, pH
7.4
NT.115
hydrophobic treated
capillaries
25 % LED, 40 % MST
NT.LabelFree
hydrophobic treated
capillaries
80 % LED, 40 % MST
7
Target Name
Ligand
MST Kd
Orthogonal Kd
Literature
Buffer
MST Conditions
NTS1 GPCR
Neurotensin
3 nM
1 nM
(43)
NT.LabelFree
standard treated capillaries
Compound
SR 48692
30 nM
52 nM
50 mM Tris pH 7.4, 50 mM NaCl,
0.1 % DDM, 0.01 % CHS, 85 mM
imidazole
Liposomes
Target Name
Ligand
MST Kd
Orthogonal Kd
Literature
Synaptotagmin-1
(in Liposome)
PtdIns(4,5) P2
Liposomes
Docking/
Tethering
Docking/
Tethering (DLS +
EM)
(44)
Synaptotagmin-1
PIP2-Liposome
36.2 ± 7.4 µM
(- Calcium)
10.6 ± 2.3 µM
(+ Calcium)
Buffer
MST Conditions
NT.115
20 mM HEPES, 150 mM KCl,
2.5 mg/ml BSA, pH 7.4
NT.115
hydrophobic treated capillaries
25 % LED, 40 % MST
20 mM HEPES, 150 mM KCl,
pH 7.4
NT.LabelFree
hydrophobic treated capillaries
80 % LED, 40 % MST
8
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11
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