Choosing mitochondrial antibodies that report the right thing: why a tom20 antibody and a tomm20 antibody are the same outer membrane import receptor under two spellings and why that makes it a mass marker rather than a function one, what an mtco2 antibody, a cox1 antibody, a cox iv antibody, an sdhb antibody and an oxphos antibody cocktail report about the respiratory chain and its two genomes, where a vdac1 antibody and a vdac2 antibody sit in the outer membrane, how an opa1 antibody, a drp1 antibody and an mff antibody report shape rather than amount, what a pink1 antibody reports only when import fails, what a tfam antibody says about the mitochondrial genome, how a ucp1 antibody and a prohibitin antibody differ from a generic mitochondria antibody or mitochondrial marker antibody, and where a citrate synthase antibody, an ogdh antibody, an ldhb antibody, a got1 antibody and a glutaminase antibody are metabolic enzymes rather than markers

Choosing mitochondrial reagents

1. Separate mass from function first. If the question is how much mitochondria a cell has, use a marker known to scale with mass. If the question is respiratory capacity, use subunits and enzyme activities and do not normalise them to each other.
2. Remember the two genomes. Respiratory complexes contain subunits encoded in the nucleus and in the mitochondrion, and they respond differently to stress and to drugs. A cocktail covering both is far more informative than any single subunit.
3. Match the reagent to the membrane. Outer membrane, inner membrane, intermembrane space and matrix proteins behave differently in fractionation and in permeabilisation experiments. Naming the sublocation is part of choosing the reagent.
4. Read shape proteins by imaging. Fusion and fission machinery levels say little about network morphology. The measurement that reports shape is imaging with a segmentation analysis, and the blot is at best supporting evidence.
5. Normalise carefully. Normalising a mitochondrial protein to a cytosolic loading control confounds mitochondrial content with expression. Where content is the variable, report both the mitochondrial marker and the total protein and let the reader see the ratio.

Mass and function pull apart

Cells adjust mitochondrial content and mitochondrial composition independently. A treatment can raise respiratory subunit levels without changing mass, or expand the network while diluting the chain. Measuring one and reporting the other is the standard confusion.

The honest presentation shows both: a mass marker and a functional readout, with the ratio stated rather than assumed.

Fractionation needs its own controls

A mitochondrial fraction contaminated with endoplasmic reticulum is common, because the two membranes are physically associated. Any fractionation claim needs markers for the contaminating compartments blotted alongside.

Show those controls in the figure. Without them a fractionation result is an assertion rather than evidence.

Common questions

Which marker is best for mitochondrial mass?

An abundant, stable structural protein rather than a respiratory subunit, because subunit levels respond to metabolic state. Confirm in your own system that the chosen marker tracks an independent measure of mass before relying on it.

Why measure subunits from both genomes?

Because nuclear and mitochondrially encoded subunits are regulated separately, and a mismatch between them is itself an informative finding about mitochondrial translation or genome maintenance.

Can a blot report mitochondrial morphology?

No. Fusion and fission protein levels are only loosely related to network shape, which has to be imaged and segmented. Reporting morphology from a blot is a claim the data does not support.

Browse by service class

• Compare Gene synthesis on advertised prices