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Membrane Transfer Buffers Solutions

Product#: Buffers-MembraneTransferBuffers
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swap_horiz Western · Southern · Northern Blotting

Membrane Transfer Buffers Solutions

This is a category overview, not an orderable page. Pick the transfer chemistry your blot needs and click View to reach its product page — the history, the selection logic, and the full formulation comparison follow below.

Membrane Transfer Buffers — Catalog · 3 Products
Select the transfer chemistry that matches your molecule and your membrane — click View for the product page.
Product Product# Core Formulation Product Page
Tris Based Transfer Buffers TrisBasedTransferBuffers Standard Tris-Glycine core Viewarrow_forward
CAPS Transfer Buffer DCP-CAPSTB0.5X 3-(Cyclohexylamino)-1-propanesulfonic acid, ~pH 11 Viewarrow_forward
Sodium Chloride and Sodium Hydroxide Transfer Buffer DCP-NACLNAOH1X Alkaline saline profile Viewarrow_forward
Looking for Bis-Tris? The formulation comparison further down covers four transfer chemistries, but only three are ordered from this page. Bis-Tris Transfer Buffer is not a separate item in this category — it is listed as FluxMPS™ Bis-Tris Transfer Buffer (DCP-BTTB1X) on the Tris Based Transfer Buffers page, alongside the Tris-CAPS and Tris-Glycine products.

Not sure which chemistry? Compare all four formulations · Match blot type to membrane · See applications · Read the FAQ

Category Snapshot

The step between the gel and the probe

Once proteins or nucleic acids are separated by size on a gel, they must be transferred to a solid support so they can be probed with antibodies or nucleic acid probes. Membrane transfer buffers are the conductive solutions that drive these macromolecules out of the gel matrix and immobilize them on a synthetic membrane (PVDF, nitrocellulose, or nylon) during Western, Southern, and Northern blotting.

  • Three products in this category, covering the standard Tris-Glycine system, a high-pH CAPS system, and an alkaline saline system for nucleic acids.
  • Four formulations are compared below — the fourth, Bis-Tris, is reached through the Tris Based Transfer Buffers page rather than ordered here.
  • Three membrane types are named by the source: PVDF, nitrocellulose, and nylon.
  • Three blot types: Western for proteins, Southern for DNA, Northern for RNA.
  • The buffer is electrochemistry, not just liquid. It has to conduct high current without generating destructive heat, set the charge profile so the target migrates uniformly, and manage gel hydration so bands do not distort.
  • pI is the variable most people forget. Most proteins carry net negative charge and migrate cleanly in standard alkaline Towbin buffer, but very basic proteins (pI above about 9) can approach neutral or reversed charge in that system.
  • Glycine is a problem for sequencing. CAPS is glycine-free, which is why it is the standard when transferred proteins will be sequenced by Edman degradation.
Western · Southern · Northern PVDF · Nitrocellulose · Nylon RUO
CATEGORY REFERENCE · MEMBRANE TRANSFER BUFFERS
The numbers behind the four transfer chemistries
  • Products in this category3
  • Formulations compared4
  • Blot types supportedWestern, Southern, Northern
  • Membrane types namedPVDF, nitrocellulose, nylon
  • Southern blot introduced1975
  • Towbin electroblotting introduced1979
  • CAPS / PVDF sequencing introduced1987
  • CAPS buffer pH~11
  • High-pI thresholdpI > ~9
  • Tris-Glycine typical protein range~15–100 kDa
Why the Transfer Buffer Matters

Moving a molecule out of a dense gel is an electrochemical problem

Moving a biomolecule out of a dense gel and onto a membrane requires a controlled electrochemical environment. Each card below is one thing the buffer has to deliver — or one way the blot fails.

bolt

Efficient electromigration

A controlled ion concentration conducts high current and voltage without excessive resistance — and without the destructive heat that excessive resistance generates.

target

Binding-site optimization

The buffer sets or preserves the charge profile of the target so it migrates uniformly toward the opposing electrode and adheres firmly to the membrane on arrival. Methanol, for example, promotes nitrocellulose binding.

straighten

Gel dimension stabilization

The buffer manages gel hydration during transfer so the matrix does not warp or shrink and distort band alignment. A distorted gel means a blot you cannot line up against your ladder.

swap_vert

High-pI accommodation

Most proteins carry net negative charge and migrate cleanly in standard alkaline Towbin buffer, but very basic proteins (pI above about 9) can approach neutral or reversed charge there. High-pH buffers such as CAPS at about pH 11 keep them moving toward the membrane.

water_drop

What methanol is actually doing

In the classic Tris-Glycine-methanol formulation, methanol counteracts gel swelling and strips SDS from proteins, improving adsorption to nitrocellulose. It is doing two jobs at once, which is why changing its concentration changes transfer behaviour.

science

Why glycine-free matters

Glycine interferes with Edman chemistry. That single fact is the reason a glycine-free CAPS system exists alongside Tris-Glycine, and why it is the buffer of choice when downstream N-terminal sequencing is the goal.

Match the buffer to the molecule, not to the protocol you inherited

The standard alkaline Towbin system is standard because it works for most proteins in the mid-range. It is not universal: a very basic protein can stall or run backwards in it, a small protein can pass straight through the membrane, and a nucleic acid needs an entirely different, alkaline saline chemistry driven by capillary flow rather than current.

~pH 11
CAPS transfer buffer — high enough to keep basic proteins net-negative and migrating
pI > ~9
the point above which a protein may approach neutral or reversed charge in standard Towbin buffer
History of Membrane Transfer Buffers

Three papers that built the blot

Each step below explains a buffer still on the shelf today — why Tris-Glycine is the default, why CAPS exists at all, and why nucleic acid transfer looks nothing like protein transfer.

  1. 1

    1975 The Southern blot

    Edwin Southern demonstrated transfer of size-separated DNA fragments from a gel onto a membrane using high-salt capillary flow, enabling sequence-specific probe detection in the “Southern blot.” This established the blotting concept for nucleic acids, but passive capillary transfer was slow and inefficient for large, tightly packed protein matrices.[1]

  2. 2

    1979 Electroblotting and the Towbin buffer

    Towbin, Staehelin & Gordon introduced electroblotting, in which an electric field pulls proteins out of a polyacrylamide gel onto nitrocellulose. Their Tris-Glycine-methanol formulation became the canonical “Towbin buffer.” Methanol counteracts gel swelling and strips SDS from proteins, improving adsorption to nitrocellulose. This invented the Western blot transfer step and set the global standard transfer chemistry still used today.[2]

  3. 3

    1987 CAPS, PVDF, and direct sequencing

    Paul Matsudaira showed proteins could be electroblotted onto PVDF membranes and sequenced directly by Edman degradation, using a glycine-free CAPS transfer buffer. This made N-terminal protein sequencing from gels routine, and established CAPS as the buffer of choice when downstream sequencing is the goal, because glycine interferes with Edman chemistry.[3]

Blot Type Reference

Which molecule, which membrane, which chemistry

The mapping the source states between the blot you are running, the molecule you are moving, the membrane it lands on, and the transfer chemistry associated with it.

Blot type Molecule transferred Membrane Transfer chemistry noted by the source
Western (wet-tank & semi-dry) Proteins, from polyacrylamide gels Nitrocellulose or PVDF Tris-Glycine-methanol (the canonical Towbin buffer); Bis-Tris for small or sensitive proteins
Southern Fragmented genomic DNA Nylon Alkaline saline, driving capillary transfer onto charge-modified nylon
Northern Single-stranded RNA Nylon Alkaline saline, driving capillary transfer onto charge-modified nylon
N-terminal sequencing Proteins, for direct Edman degradation PVDF CAPS at about pH 11, glycine-free
Protein transfer and nucleic acid transfer are not the same operation. Western blotting is driven by an electric field pulling proteins out of the gel. Southern and Northern blotting, in the system this category covers, are driven by capillary flow through an alkaline saline buffer that simultaneously denatures DNA duplexes. Choosing between them is a choice of physics, not just of reagent.
Applications

What you are transferring, and why

Select a workflow to see what the source says about it, and which formulation it names for that job.

Proteins · nitrocellulose or PVDF
  • Western blotting (wet-tank & semi-dry) — driving proteins from polyacrylamide gels onto nitrocellulose or PVDF
  • Named formulation: Tris-Based Transfer Buffers, the universal Western blot standard for routine wet-tank transfer of mid-range proteins (~15–100 kDa), typically paired with user-added methanol to optimize nitrocellulose adhesion
Nucleic acids · nylon · capillary transfer
  • Southern & Northern blotting — transferring fragmented genomic DNA or single-stranded RNA onto nylon membranes
  • Named formulation: NaCl / NaOH Transfer Buffer, optimized for capillary blotting of nucleic acids — the alkaline environment simultaneously denatures DNA duplexes and drives capillary transfer of target sequences onto charge-modified nylon membranes
Proteins · PVDF · Edman degradation
  • N-terminal protein sequencing — CAPS transfer onto PVDF for direct Edman degradation
  • Named formulation: CAPS Transfer Buffer — high pH keeps basic proteins net-negative and migrating, and it is glycine-free, making it the standard when transferred proteins will be sequenced by Edman degradation
Large proteins that transfer poorly
  • High-molecular-weight protein transfer — customized buffer/methanol conditions to mobilize large proteins that transfer poorly
  • The source names no specific product for this workflow; it identifies the buffer and methanol conditions as the variables to customize. For a custom formulation, contact support@diagnocine.com.
Complex protein arrays
  • Two-dimensional (2D) gel transfer — immobilizing complex protein arrays for downstream immunodetection or mass spectrometry
  • The source names no specific product for this workflow.
Formulation Comparison

All four transfer chemistries side by side

The source comparison table, reproduced row for row. Three of these four are ordered from the catalog at the top of this page; Bis-Tris is reached through the Tris Based Transfer Buffers page.

Product Core Formulation Best Use
Tris-Based Transfer Buffers Standard Tris-Glycine core Universal Western blot standard — routine wet-tank transfer of mid-range proteins (~15–100 kDa); typically paired with user-added methanol to optimize nitrocellulose adhesion
Bis-Tris Transfer Buffer Bis-Tris core, milder near-neutral pH Optimized for small/sensitive proteins — a less alkaline environment that helps retain low-molecular-weight proteins (<~20 kDa) that can blow through the membrane in harsher systems
CAPS Transfer Buffer 3-(Cyclohexylamino)-1-propanesulfonic acid, ~pH 11 Choice for high-pI proteins & N-terminal sequencing — high pH keeps basic proteins net-negative and migrating; glycine-free, making it the standard when transferred proteins will be sequenced by Edman degradation
NaCl / NaOH Transfer Buffer Alkaline saline profile Optimized for capillary blotting of nucleic acids — the alkaline environment simultaneously denatures DNA duplexes and drives capillary transfer of target sequences onto charge-modified nylon membranes
Two of these four names differ from the published product names. The row labelled NaCl / NaOH Transfer Buffer is sold as Sodium Chloride and Sodium Hydroxide Transfer Buffer (DCP-NACLNAOH1X), and the row labelled Bis-Tris Transfer Buffer is listed as FluxMPS™ Bis-Tris Transfer Buffer (DCP-BTTB1X) on the Tris Based Transfer Buffers page. Both table labels and both product names are reproduced exactly as published; neither has been edited.
FAQ

Frequently asked questions

The questions that come up most often when a blot does not look the way it should.

Four things. It conducts high current and voltage without excessive resistance and the destructive heat that follows. It sets or preserves the charge profile of the target so it migrates uniformly and adheres firmly to the membrane on arrival. It manages gel hydration so the matrix does not warp or shrink and distort band alignment. And in high-pH systems it keeps very basic proteins net-negative so they keep moving toward the membrane.
Tris-Based Transfer Buffers, on the standard Tris-Glycine core. The source calls it the universal Western blot standard for routine wet-tank transfer of mid-range proteins, roughly 15 to 100 kDa, typically paired with user-added methanol to optimize nitrocellulose adhesion. This is the direct descendant of the 1979 Towbin formulation.
Move to a high-pH system. Most proteins carry net negative charge and migrate cleanly in standard alkaline Towbin buffer, but very basic proteins with a pI above about 9 can approach neutral or even reversed charge there — which is exactly what a protein that will not move, or moves the wrong way, looks like. CAPS Transfer Buffer at about pH 11 keeps them net-negative and migrating toward the membrane.
Yes, and this is the one case where the buffer choice is not negotiable. Glycine interferes with Edman chemistry, so a glycine-free system is required. CAPS Transfer Buffer is glycine-free, which is why it became the standard when transferred proteins will be sequenced by Edman degradation. The method comes from Matsudaira in 1987, who showed proteins could be electroblotted onto PVDF and sequenced directly.
Sodium Chloride and Sodium Hydroxide Transfer Buffer (DCP-NACLNAOH1X), listed in the comparison table as the NaCl / NaOH Transfer Buffer. It is optimized for capillary blotting of nucleic acids: the alkaline environment simultaneously denatures DNA duplexes and drives capillary transfer of target sequences onto charge-modified nylon membranes. Southern blotting transfers fragmented genomic DNA and Northern blotting transfers single-stranded RNA, both onto nylon.
Bis-Tris Transfer Buffer, on a Bis-Tris core with a milder near-neutral pH. The source describes it as optimized for small and sensitive proteins — a less alkaline environment that helps retain low-molecular-weight proteins below roughly 20 kDa that can blow through the membrane in harsher systems. It is not a separate item in this category: it is listed as FluxMPS™ Bis-Tris Transfer Buffer (DCP-BTTB1X) on the Tris Based Transfer Buffers page.
Because methanol is doing two jobs at once in the classic Tris-Glycine formulation: it counteracts gel swelling and it strips SDS from proteins, which improves adsorption to nitrocellulose. That is also why it appears as a variable rather than a fixed setting — for high-molecular-weight proteins that transfer poorly, the source names customized buffer and methanol conditions as the way to mobilize them. It publishes no target percentage, so treat it as something to titrate for your target rather than a number to copy.
Key References

The three papers behind the category

The source's own reference list, verified against the published record.

  1. Southern, E. M. (1975). Detection of specific sequences among DNA fragments separated by gel electrophoresis. Journal of Molecular Biology, 98(3), 503–517. — The origin of blotting: capillary transfer of DNA to a membrane for sequence-specific detection (the “Southern blot”).doi:10.1016/S0022-2836(75)80083-0
  2. Towbin, H., Staehelin, T., & Gordon, J. (1979). Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proceedings of the National Academy of Sciences, 76(9), 4350–4354. — The paper that invented Western blot electroblotting and defined standard Tris-Glycine transfer chemistry.doi:10.1073/pnas.76.9.4350
  3. Matsudaira, P. (1987). Sequence from picomole quantities of proteins electroblotted onto polyvinylidene difluoride membranes. Journal of Biological Chemistry, 262(21), 10035–10038. — Introduced PVDF electroblotting and the glycine-free CAPS transfer system for direct protein sequencing.
Transfer buffer selection support. For help matching a chemistry to a specific molecule, membrane, or blotting system, for customized buffer and methanol conditions, or for documentation requests, contact support@diagnocine.com. Ready to order? Back to the Membrane Transfer Buffers catalog.

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