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A whey protein used as a model solute in studies of protein association and transport. Its effective electric charge, association state and hydrodynamic radius depend on solution conditions. In a specified spherical-particle model, its electrophoretic mobility and sedimentation speed depend on its charge and buoyant mass through the same Stokes drag law.

Diffusion and mobility. For a dilute isolated sphere in a Newtonian fluid, Stokes's law gives the drag magnitude $F_{\mathrm{drag}}=6\pi \eta Rv=\xi v$, where $\eta$ is the dynamic viscosity and $R$ is the hydrodynamic radius. The mechanical mobility is therefore $1/\xi$; the translational hydrodynamic mobility matrix is ${\xi }^{-1}I$.

To relate mobility to diffusion, put the particles in a weak external potential energy $U(x)$. Their drift speed is $-U^{\prime }(x)/\xi$. For number density $n(x)$, the sum of drift and Fick's first law fluxes isAt thermal equilibrium, the Boltzmann distribution gives $n\propto e^{-U/(k_{B}T)}$, and the net flux is zero. Substituting $n^{\prime }=-n{U}^{\prime }/(k_{B}T)$ into $J=0$ yields $D=k_{B}T/\xi$. Thus the Stokes–Einstein relation isHere $k_B$ is the Boltzmann constant and $T$ is absolute temperature. The assumptions include a no-slip approximately spherical particle, a continuum fluid, a small particle Reynolds number, and negligible hydrodynamic interactions with other particles or boundaries.

Gravitational sedimentation. The buoyant force must be included, since the mass of displaced water is comparable to the protein mass. Using $1\mathrm{Da}=1.66054\times 10^{-27}\mathrm{kg}$ givesso the buoyant mass in the stated spherical model isThe drag coefficient is $\xi =6\pi (10^{-3})(2.16\times 10^{-9})=4.0715\times 10^{-11}\mathrm{kg}{\mathrm{s}}^{-1}$. At terminal speed, $\xi v_g=m_{b}g$, soIgnoring buoyancy would give $1.47\times 10^{-11}\mathrm{m}{\mathrm{s}}^{-1}$ and would not use the supplied water density. This calculation treats the stated radius as both the hydrodynamic radius and the radius determining displaced volume; a real hydrated protein can require a separately measured partial specific volume.

Electrical drift. In the specified simple electrophoresis model, terminal force balance is $Ze\mathbf{E}=\xi \mathbf{v}$. Thereforewhere $\mu$ is the electrophoretic mobility and $e$ the elementary electric charge. For the given positive beta-lactoglobulin charge,in the direction of the electric field. This exceeds the gravitational sedimentation speed by about $8.6\times 10^6$.

Separation time. The two populations have equal drag coefficients because their radii are equal. Their difference in drift speed is therefore set by one elementary charge:Starting with coincident band centres, their separation is $\Delta vt$. HenceAt that time the original band has travelled $20\mathrm{mm}$ and the mutant $21\mathrm{mm}$. Their masses do not enter this ideal electrical force balance: equal-radius particles with different masses but the same charge would have equal electrophoretic mobility in this model.

Practical discrimination. A one-charge difference is enough for the stipulated separation, so the method is sensitive to mutations that change effective charge or friction. It cannot distinguish every mutation: substitutions that preserve charge and hydrodynamic size need not separate. Buffer pH changes protein ionization, and realistic electrophoresis also involves the ionic atmosphere, electro-osmosis, walls or gel interactions, and finite band widths.

For example, the Stokes–Einstein relation at $293.15\mathrm{K}$ gives $D=9.94\times 10^{-11}{\mathrm{m}}^2{\mathrm{s}}^{-1}$. For an initially narrow band, diffusive broadening of electrophoretic bands givesat the calculated separation time. Thus the centres are about $3.15$ diffusion standard deviations apart, rather than forming infinitely sharp bands. The stated $1\mathrm{mm}$ centre-separation criterion is achievable in the model, but actual resolution also depends on the loading width and other dispersion mechanisms.

Equivalent centrifugal acceleration. For the original protein to attain its electrical drift speed by sedimentation, its buoyant centrifugal force must equal $20eE$:ConsequentlyThis is the acceleration at the protein's position in the rotating solution; no rotation rate can be assigned without a centrifuge radius. If the intended comparison is instead the relative separation speed of the two bands, use $\Delta m_b=600\mathrm{Da}$, since their displaced volumes are equal. Matching $|\Delta m_b|a_{\mathrm{eff}}=eE$ then gives $8.04\times 10^7\mathrm{m}{\mathrm{s}}^{-2}$. In that separation experiment the heavier original protein moves faster centrifugally, whereas the more highly charged mutant moves faster electrically.

A colloidal grain in a liquid is large enough to see with a microscope, yet small enough for thermal motion to compete with its tendency to settle. In the Perrin sedimentation experiment, the equilibrium number density falls with height: a visible miniature atmosphere. Counting grains at different heights measures an energy scale; tracking their Brownian motion measures a transport rate. Connecting both to the same Avogadro constant makes the experiment a quantitative test of kinetic theory. The Wiener measure describes an idealized path law; these measurements instead test a physical model and its material parameters.

Jean Baptiste Perrin's 1909 memoir traces his sedimentation experiments to May 1908. It reports relative counts $100$, $47$, $22.6$, $12$ at heights $5$, $35$, $65$, $95\mu \mathrm{m}$, and compares radius estimates from Stokes settling velocity, counted grain mass, and rows of touching grains.^[ref]

The 1926 lecture describes resin precipitated from alcohol into water, washed and fractionated by centrifugation to obtain nearly monodisperse gamboge or mastic grains. Diameter and density were measured separately. A sealed slide-and-cover-glass chamber prevented evaporation; a vertical microscope resolved roughly $2\mu \mathrm{m}$ layers through a roughly $0.1\mathrm{mm}$ depth. Refocusing and photographing compared layer populations after equilibrium; changing temperature reversibly changed the distribution.^[ref]

In Les Atomes (1913), Jean Baptiste Perrin restricts the eyepiece view to fewer than five or six grains and alternates observations between heights. Focus displacement requires a refractive index correction to recover actual layer separation. Horizontal positions were marked with a camera lucida, usually every $30\mathrm{s}$, and distances calibrated with a micrometer; the polygon joins sampled positions rather than recording the intervening trajectory.^[ref][ref]