MACS Matchmaker
The lino anti-tag adapter system uses oriented capture proteins (Protein A/G + Protein A/G/L, trisNTA, Strep-Tactin XT, anti-GFP VHH) that are pre-conjugated to a chosen oligonucleotide sequence and ship ready-to-use from the catalogue. Each adapter hybridizes to the Oligo|PEG chip via DDI, then the tagged ligand is loaded in a capture step. The chip is regenerable at the duplex level between different tagged ligands, and the captured ligand can additionally be eluted via its tag (e.g. imidazole for His-tag, low pH for Fc-captured IgGs) without stripping the duplex.
Before you start
- Read Safety & Compliance and the safety data sheet for every reagent below. The SDS, not this page, governs how a reagent is handled, stored and disposed of.
- Use the personal protective equipment and the waste route your laboratory prescribes for these reagent classes.
- Check this protocol against the materials actually in hand: confirm the intended Seq ID, the product revision, the lot-specific certificate and the instructions supplied with it. Where those differ from this page, the documentation supplied with the material governs.
Available adapters
All catalogue adapters share the same format: 110 µL stock at 2.6 µM in PBST, enough for 10 immobilizations at the recommended 200 nM working concentration (130 µL/injection). Each is pre-conjugated to a single Seq ID (01–64) chosen at order time; Seq ID 00 is reserved for the backfilling strand — combine adapters with different Seq IDs to multiplex on a standard 8×8 Oligo|PEG chip (6×9 on request).
| Adapter | Tag / ligand captured | Elution | Webshop |
|---|---|---|---|
| Protein A/G + Protein A/G/L | Human / mouse IgG (Fc region) for the A/G variant; A/G/L also captures kappa light chains (Fabs and scFvs with a κ chain) | pH 2.0 glycine or duplex strip | View in shop |
| TrisNTA | 6×His-tag (requires NiSO₄ pre-loading — not included) | 250 mM imidazole or 50 mM EDTA | View in shop |
| Strep-Tactin XT | Strep-tag II and Twin-Strep-tag | 5 mM desthiobiotin or duplex strip | View in shop |
| Anti-GFP VHH | GFP-fusion proteins | Duplex strip | View in shop |
General protocol (all adapters)
Each adapter follows the same three-step DDI capture workflow:
- Adapter-oligo hybridization. Flow the adapter-oligo conjugate (200 nM in PBST) over the Oligo|PEG chip for 5 min. The chip is now loaded with oriented capture sites.
- Tagged ligand capture. Flow the tagged protein at a sub-saturating concentration (typically 10–100 nM) for 3–5 min. Monitor the signal in real time and stop when the desired loading level is reached.
- Analyte measurement. Run the analyte concentration series at 30 µL/min (6-ch) / 60 µL/min (3–4-ch) / 100 µL/min (1-ch) for association and 200 µL/min for dissociation. Regenerate with tag-specific elution or strip the duplex for a fresh adapter load.
Protein A/G — IgG and antibody fragment capture
| Parameter | Value |
|---|---|
| Adapter | Protein A/G-oligo (Fc capture) or Protein A/G/L-oligo (Fc + Vκ capture) — order one Seq ID per loaded ridge |
| Adapter stock | 110 µL at 2.6 µM in PBST (10 immobilizations per vial) |
| Working dilution | 200 nM in PBST, 130 µL injection |
| Hybridisation flow rate / contact time | 10 µL/min, 5 min |
| IgG loading concentration | 10–100 nM in PBST |
| Loading flow rate / contact time | 10 µL/min, 3–5 min (to desired signal, not necessarily saturation) |
| SCK association flow rate | 30 µL/min (6-ch) / 60 µL/min (3–4-ch) / 100 µL/min (1-ch) |
| SCK dissociation flow rate | 200 µL/min |
| Regeneration option A (tag elution) | 10 mM glycine pH 2.0, 30 s — releases IgG, leaves adapter on chip |
| Regeneration option B (duplex strip) | 3 M GuHCl + 125 mM NaOH, 400 µL/min, 0.5 min |
TrisNTA — His-tag capture
| Parameter | Value |
|---|---|
| Adapter | TrisNTA-oligo, one Seq ID per loaded ridge |
| Adapter stock | 110 µL at 2.6 µM in PBST (10 immobilizations per vial) |
| Working dilution | 200 nM in PBST, 130 µL injection |
| Hybridisation flow rate / contact time | 10 µL/min, 5 min |
| Ni²⁺ pre-loading | 500 µM NiSO₄ in PBST (NiSO₄ not included in the adapter vial), 10 µL/min, 2 min; wash with PBST (200 µL/min) to remove excess Ni²⁺ before loading His-tagged protein |
| His-tagged ligand concentration | 10–100 nM in PBST |
| Loading flow rate / contact time | 10 µL/min, 3–5 min |
| SCK association flow rate | 30 µL/min (6-ch) / 60 µL/min (3–4-ch) / 100 µL/min (1-ch) |
| SCK dissociation flow rate | 200 µL/min |
| Regeneration option A (His elution) | 250 mM imidazole in PBST — releases His-tagged protein |
| Regeneration option B (chelation) | 50 mM EDTA in PBST — strips Ni²⁺ and ligand; re-load Ni²⁺ to reuse |
| Regeneration option C (duplex strip) | 3 M GuHCl + 125 mM NaOH, 400 µL/min, 0.5 min |
Strep-Tactin XT — Strep-tag II capture
| Parameter | Value |
|---|---|
| Adapter | Strep-Tactin XT-oligo, one Seq ID per loaded ridge |
| Adapter stock | 110 µL at 2.6 µM in PBST (10 immobilizations per vial) |
| Working dilution | 200 nM in PBST, 130 µL injection |
| Hybridisation flow rate / contact time | 10 µL/min, 5 min |
| Strep-tagged ligand concentration | 10–50 nM in PBST |
| Loading flow rate / contact time | 10 µL/min, 3–5 min |
| SCK association flow rate | 30 µL/min (6-ch) / 60 µL/min (3–4-ch) / 100 µL/min (1-ch) |
| SCK dissociation flow rate | 200 µL/min |
| Regeneration (tag elution) | 5 mM desthiobiotin in PBST, 2 min — releases Strep-tag II ligand; Strep-Tactin XT adapter remains on chip for next capture |
| Regeneration (duplex strip) | 3 M GuHCl + 125 mM NaOH, 400 µL/min, 0.5 min |
QC and acceptance
Capture is two hybridization-and-binding steps in series, and they fail for unrelated reasons. Record the response of each step separately — the pair identifies which one failed, where a single end-point measurement cannot.
- Adapter loading. The adapter injection gives a response that persists through the wash. No response here is a loading failure: wrong or degraded adapter, a surface that was not available, or a buffer that suppresses hybridization. Nothing downstream can succeed, so stop and fix it.
- Ligand capture. With the adapter loaded, the ligand injection gives its own persistent response. Loading that worked followed by capture that did not points at the ligand rather than the surface — tag absent, cleaved or sterically occluded, a mismatched adapter, or a pH at which that adapter does not bind its tag.
- Capture stability. Affinity capture is reversible by construction. Watch the baseline between loading and analyte injection: a steady downward drift is ligand leaving the adapter, and it subtracts from every dissociation measured afterwards.
Practical notes
See also: immobilization method comparison table.