One-step affinity purification using ALFA Selector^ST magnetic. A, Purified GFP-ALFA (left lane) was blended into HeLa lysate (Input) and incubated with ALFA Selector^ST magnetic. After washing, the resin was successively incubated with 500 μM ALFA peptide for 2x 10 min (E1: Peptide) before eluting remaining protein with SDS sample buffer (E2: SDS). Indicated fractions were analyzed by SDS–PAGE and Coomassie staining. Eluate fractions correspond to the material eluted from 1 μL of resin. B, control experiment without substrate.

ALFA Selector ST

Name: ALFA Selector ST
SKU: N1511
Availability: InStock

Cat No: N1511 Category: Selector Resins

275,00 € – 445,00 €

ALFA Selector ST (for “Super Tight”) is an innovative, high-affinity resin (Agarose & Magnetic) designed for efficient and clean pull-down of ALFA-tagged proteins.

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ALFA Selector ST (for “Super Tight”) is an innovative, high-affinity resin designed for the highly efficient pull-down of ALFA-tagged proteins.

Please look to our Technology Section for more information about the ALFA system.

ALFA Selector ST is based on a single-domain antibody (sdAb), which is covalently immobilized on 4% cross-linked agarose or agarose-magnetic beads and binds ALFA-tagged proteins with extraordinarily high affinity (Kd = 26 pM). Target proteins are efficiently released from ALFA Selector ST under acidic (e.g., 0.1 M glycin pH2.2) or denaturing conditions. The extremely tight interaction with ALFA-tagged proteins makes competitive elution using ALFA elution peptide inefficient. We recommend using ALFA Selector PE for competitive elution under physiological and room temperature condition conditions. For efficient elutions at 4°C, we recommend using the ALFA Selector CE

The innovative, oriented and highly selective attachment of the active sdAbs via a flexible linker guarantees optimal accessibility for target proteins and at the same time largely eliminates batch-to-batch variations. Due to the single-chain nature of sdAbs and their covalent attachment, no “leakage” of light and heavy chains from IgGs is observed even during elution with strongly denaturing and reducing conditions (e.g., SDS sample buffer). ALFA Selector ST thus features high affinity and superior capacity for ALFA fusion proteins while showing negligible non-specific background.

ALFA Selector ST is compatible not only with physiological buffers but also with high stringency buffers (see ALFA Selector Manual). ALFA Selector ST thus provides great freedom to adjust the binding and washing conditions to the experimental needs.

MSDS

Variations:

Conjugation	Amount	Cat No.	RRID
4% cross-linked magnetic agarose	2000 μl	N1516-L	AB_3075994
4% cross-linked agarose	2000 μl	N1511-L	AB_3075991

ALFA Selector ST

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