What does a western blot diagram show?

It shows the method: sample prep, SDS-PAGE, transfer to a membrane, blocking, primary and secondary antibody incubation, and detection. It does not show results. Real data is the blot image from your own experiment.

What is the order of the western blot transfer sandwich?

From the cathode (-) to the anode (+): sponge, filter paper, gel, membrane, filter paper, sponge. The gel is on the cathode side and the membrane on the anode side, so proteins move from the gel to the membrane. Semi-dry systems use filter paper stacks instead of sponges.

Can I use this to make a western blot figure for a paper?

You can use it for the method panel, such as a workflow or antibody schematic. It cannot replace the blot itself. Do not draw or generate bands, and show real blots with molecular weight markers and a loading control as your target journal requires.

What is the difference between wet and semi-dry transfer?

Wet transfer submerges the sandwich in a buffer tank between sponges. Semi-dry transfer presses buffer-soaked filter paper, gel and membrane directly between plate electrodes. Switch between them in the generator to see both stacks.

Why is the secondary antibody enzyme-linked?

The primary antibody recognizes the target but is unlabeled. The secondary antibody recognizes the primary and carries an enzyme such as HRP, which turns the substrate into light. One primary can carry several secondaries, which amplifies the signal.

Is the western blot diagram generator free?

The precise diagram tab is free to use and to download as SVG or PNG, with no sign-up. Sign in only if you want AI illustrations or other ConceptViz tools.

Biology Tool

Western Blot Diagram Generator method schematic

Draw a western blot workflow, transfer sandwich, or antibody detection schematic. Edit labels, pick wet or semi-dry, export SVG or PNG. Method figures only.

Six-step workflow with editable labelsTransfer sandwich: wet or semi-dryPrimary and HRP secondary antibody schematicDownload SVG or PNG, free

Method schematic as SVG or PNG: workflow, transfer sandwich, antibody detection, or a blank worksheet. Free, no account.

Figure options

Method schematic only. Never use it as, or in place of, real blot data.

Transfer type

Title

Primary antibodySecondary antibody

Step labels

Western Blot Diagram Examples

Four exact diagrams from the Precise Diagram tab and two AI examples with their prompts and flaws. All are method schematics, not data.

View:

Labeled workflow

Exact diagram from the Precise Diagram tab. Dashed outlines in the gel are schematic, not data.

workflowlabeled

Transfer sandwich

Exact diagram. Gel sits on the cathode (-) side, membrane on the anode (+) side.

transfersandwich

Antibody detection

Exact diagram. Primary binds the target, the HRP secondary binds the primary.

antibodydetection

Blank worksheet

Exact diagram. Same six panels with the step names removed for revision.

blankworksheet

AI example: workflow

AI illustration, checked by hand after one prompt revision. Electrodes, wells and antibody orientation are correct. Flaws: the transfer step is an exploded view with an arrow but no cathode or anode labels; the gel is drawn inside a thick tank frame; crowded small text.

aiworkflow

AI example: transfer sandwich

AI illustration, checked by hand. Layer order and electrodes are correct. Flaws: exploded view with gaps between layers, the arrow sits beside the stack instead of through it, the callout text hugs the left edge, and the membrane is drawn as a thick slab.

aisandwich

What a western blot diagram shows

A western blot diagram is a method figure: how proteins are separated on a gel, moved to a membrane, and detected with antibodies. It explains the procedure. It is not a result. This page draws the procedure as an exact diagram, and every membrane or gel shape in it is an outlined schematic with no band images.

The six steps in order

1. Sample prep: lyse cells or tissue and denature proteins in SDS sample buffer.
2. SDS-PAGE: SDS gives proteins a negative charge, so they separate mainly by size as they run toward the anode (+).
3. Transfer: an electric field moves proteins from the gel onto a nitrocellulose or PVDF membrane. Wet (tank) and semi-dry systems are both common.
4. Blocking: a protein solution such as milk or BSA covers empty membrane sites so antibodies do not stick everywhere.
5. Antibodies: an unlabeled primary antibody binds the target, then an enzyme-conjugated secondary (often HRP) binds the primary.
6. Detection: HRP acts on a chemiluminescent substrate, and the light is captured on film or with a digital imager.

The transfer sandwich order

From the cathode (-) to the anode (+): sponge (fiber pad), filter paper, gel, membrane, filter paper, sponge. The gel faces the cathode and the membrane faces the anode, because SDS-coated proteins are negative and move toward the positive electrode.
Semi-dry transfer drops the sponges and stacks buffer-soaked filter paper between plate electrodes. The gel-to-membrane direction stays the same. Sponge and paper counts differ by apparatus, so follow your manufacturer manual.

Common drawing mistakes

Sandwich reversed: membrane on the cathode side. Proteins would run off the gel away from the membrane.
Primary and secondary swapped: the secondary carries the enzyme and binds the primary, not the target protein.
Enzyme drawn on the primary antibody in an indirect scheme, or substrate drawn before the secondary step.
Missing filter paper, or a membrane placed outside the gel footprint.
Bands drawn as if they were results. A method figure should never include band images.

Integrity note: schematics are not data

These diagrams are method illustrations. Never use them as, or in place of, real blot data, and never edit a method figure to look like a blot. Real blot figures should show the uncropped membrane or gel with molecular weight markers and a loading control, as your journal requires. AI illustrations can be wrong in small ways, so check every label against the exact diagram and discard any image that looks like a real blot.

Precise diagram vs AI illustration

The Precise Diagram tab fixes the layer order and labels in code, lets you edit the six step labels, switch wet or semi-dry, recolor the antibodies, and export SVG or PNG. The AI Illustration tab is for slide mockups; its prompts ask for method schematics only.

Frequently Asked Questions

Sources checked

Workflow order and detection steps follow the Thermo Fisher western blotting overview. Gel-to-cathode and membrane-to-anode orientation follows the QIAGEN western blotting bench guide. Checked October 2026. Your apparatus manual decides sponge and paper counts.