Blotto — Western blot quantification

The problem

Quantification is the part no one can check.

In most labs, blot analysis is taught hand to hand — one trainee to the next. Each pass tends to lose a detail. The PI sees finished figures and has to trust the analysis underneath them. It holds up right until a reviewer asks how a band was measured — and there's no record to point to.

Two people ran the same blot and got different numbers. The afternoon went to working out why.

A reviewer asked how you handled background and saturation. The methods section didn't really answer it.

A new student needed three weeks to learn a quantification workflow that was never fully written down.

How it works

One path, from images to defensible numbers.

You decide where to look and what the result means. Blotto does the measuring — the same way, every time — and shows its work.

Load your exposure series

Drop in the bracketed exposures of a single blot. Blotto reads the acquisition metadata — exposure time, sensor ceiling, dimensions — directly from the file rather than guessing at it.

Confirm lanes and bands

Automatic lane and band detection that you review and correct, with an editable lane count and outer-lane anchoring. You target; Blotto measures. Nothing is finalized without your confirmation.

Slope across the linear range

For each band, Blotto fits intensity against exposure duration, keeps only the linear range, and excludes points that saturate against the sensor ceiling. The slope is the quantity — and it stays comparable across strong and weak bands on the same blot, where no single exposure captures both well.

Export with the calculation shown

Per-band numbers as CSV, alongside the chart that shows the fit, the range used, and the points it left out. If a reviewer asks how you measured, you hand them the file.

export · per-band, with the steps that produced it

lane,band,slope,linear_range,points_used,saturation
1,1,  142.7,  exp 1–4,   4,        ok
1,2,   38.4,  exp 3–6,   4,        ok
2,1,  291.0,  exp 1–3,   3,        2 excluded (ceiling)

Why it holds up

Standardized, and shown — not asserted.

Blotto's measurement is anchored to recognized densitometry — rolling-ball background subtraction and linear-range slope fitting — computed on the full-bit-depth data, never the image on your screen. The method runs identically for every analyst and every blot, and every number arrives with the steps that produced it.

That's the difference between a number a reviewer has to trust and one they can check.

Plainly

What Blotto is — and isn't.

What it isn't

—A replacement for your imager or for ImageJ.

—A normalization tool. Blotto gives you per-band numbers; how you normalize is your analysis.

—A fully automatic pipeline. You confirm every lane and band.

—Validated for clinical or diagnostic use.

What it is

+A standardization and documentation layer for the measurement itself.

+The same method, every analyst, every blot — measured on the true data and shown plainly.

+Output that's consistent, comparable, and defensible — without adding hours.

+Browser-based. Your images stay on your machine.

Where we are

We're validating accuracy before we claim it.

A more elaborate method can be more wrong while looking more rigorous. The only honest test is to measure known ratios — a dilution series at 1×, 2×, 4×, 8× — and check that the tool recovers them.

We're running that validation now, with a small number of independent labs. Until it's done, we make no best-in-class accuracy claim. What's real today is standardization, transparency, and a defensible record — worth having on their own, whatever the accuracy work returns.

Run your blots the right way — with a record that holds up.

Early access is free and opens lab by lab. Request a code; when your slot opens you'll have 14 days of full access to run your own blots.

Got it — we'll be in touch personally. You can also reach us at nathan@leftbrainconsult.com.

We reach out personally. No automated sequences.