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For a poster with detailed information on the ALFA System and its applications click here.  The ALFA-tag is a novel, rationally designed epitope tag (SRLEEELRRRLTE) that forms a small and stable α-helix that is functional irrespective of its position on the target protein in prokaryotic and eukaryotic hosts. The flanking proline residues are not part of the recognition sequence, but may aid in spacial separation of the tag from the protein of interest.  ALFA-tag for imaging  The ALFA-tag has the broadest spectrum of applications in life sciences compared to any other tag. We developed a nanobody binding the ALFA-tag with ~26 pM affinity, which is ideally suited not only for super-resolution microscopy, immunoprecipitations and Western blotting, but also allows detection of proteins within living cells. This nanobody can be conjugated to vitually all kinds of fluorophores, enyzmes or other biomolecules. Check out the FluoTag-X2 anti-ALFA in our webshop.  ALFA-tag for immunoprecipitations  For immunoprecipitations we offer two types of ALFA Selector resins. Both ALFA Selectors are based on covalently coupled nanobodies that specifically recognize ALFA-tagged targets. ALFA Selector ST (for Super-Tight) offers the highest possible affinity for ALFA-tagged targets, which can be eluted under acidic or denaturing conditions. ALFA Selector PE (for Peptide Elution) displays an engineered nanobody with lower affinity (Kd ~11 nM), which is optimized for peptide elution under physiological conditions. Both Selector resins feature low non-specific protein adsorption and therefore allowing clean and meaningful immunoprecipitations. Due to the covalent and oriented immobilization, the immobilized nanobodies will stay on the resin even when using harsh denaturing and/or reducing conditions for elution. In contrast to conventional immunoprecipitations, eluates from both ALFA Selectors will therefore be free from large amounts of antibody fragments. HeLa mock lysate perpared in PBS was mixed with recombinant purified ALFA-tagged GFP substrate (right lanes). The lysate/substrate mixture was incubated with either ALFA SelectorPE (left panel) or ALFA SelectorST (right panel). After washing with PBS, bound proteins were eluted either with 200µM ALFA-peptide, glycin pH2.2 or SDS sample buffer (SDS). Peptide elution under native conditions is efficient when using ALFA SelectorPE. The target protein can be recovered from both ALFA Selector resins by elution with glycin or SDS sample buffer.  Note that the purity of the target protein eluted from both ALFA Selector resins using low pH or peptide (only ALFA SelectorPE) is superior to the input material, which has been purified by a 2-step chromatography process involving affinity chromatography with proteolytic release and gel filtration.   Related ALFA products in our webshop FluoTag-X2 anti-ALFA                 ALFA Selector ST             ALFA Selector PE incl. ALFA elution peptide               ALFA elution peptide

  • ALFA elution peptide

    ALFA elution peptide

    One vial contains 10 mg of ALFA elution peptide to prepare a native elution buffer for the ALFA Selector PE. We...

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  • ALFA Selector ST

    ALFA Selector ST

    ALFA Selector ST (for "super tight") is an innovative, high affinity resin designed for the highly efficient pull-down of ALFA-tagged...

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  • ALFA Selector PE incl. ALFA elution peptide

    ALFA Selector PE incl. ALFA elution peptide

    ALFA Selector PE (for "peptide elutable") is an innovative affinity resin designed for the highly efficient purification of native ALFA-tagged...

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  • FluoTag-X2 anti-ALFA

    FluoTag-X2 anti-ALFA

    FluoTag®-X2 anti-ALFA is derived from an in-house developed single-domain antibody (sdAb) that recognize a small rationally-designed epitope tag: the...

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  • FluoTag®-X2 anti-VGLUT1

    FluoTag®-X2 anti-VGLUT1

    FluoTag®-X2 anti-VGLUT1 is derived from a single-domain antibody (sdAb) recognizing specifically VGLUT1 with high affinity and specificity. Our FluoTag-X2...

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  • FluoTag®-X2 anti-CD62L

    FluoTag®-X2 anti-CD62L

    FluoTag®-X2 anti-CD62L is derived from an in-house developed single-domain antibody (sdAb) recognizing the human lymphocyte marker CD62L ...

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  • FluoTag®-X2 anti-Rabbit IgG

    FluoTag®-X2 anti-Rabbit IgG

    FluoTag®-X2 anti-Rabbit is an isotype- and species-specific FluoTag® directed against immunoglobulins from Rabbit. In our FluoTag-X2 series, two fluorophore...

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  • FluoTag®-X2 anti-Mouse IgG

    FluoTag®-X2 anti-Mouse IgG

    FluoTag®-X2 anti-Mouse is a species-specific FluoTag® directed against the kappa light chain from Mouse. In our FluoTag-X2 series, two...

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  • FluoTag®-X2 anti-mScarlet-i

    FluoTag®-X2 anti-mScarlet-i

    FluoTag®-X2 anti-mScarlet-i is derived from an in-house developed single-domain antibody (sdAb) recognizing mScarlet-i with high affinity and specificity. Our...

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  • FluoTag®-X2 anti-Guinea Pig IgG

    FluoTag®-X2 anti-Guinea Pig IgG

    FluoTag®-X2 anti-Guinea Pig is an isotype- and species-specific FluoTag® directed against immunoglobulins from Guinea Pig. The FluoTag-X2 anti-Guinea Pig...

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  • FluoTag®-X2 anti-Chicken IgY

    FluoTag®-X2 anti-Chicken IgY

    FluoTag®-X2 anti-Chicken is an isotype- and species-specific FluoTag® directed against immunoglobulins from Chicken. In our FluoTag-X2 series, two fluorophore...

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  • GFP Selector

    GFP Selector

    GFP Selector is based on a high-affinity single-domain antibody (sdAb) that is covalently immobilized on 4% cross-linked agarose beads. The innovative,...

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  • RFP Selector

    RFP Selector

    RFP Selector is based on a high-affinity single-domain antibody (sdAb) that is covalently immobilized on 4% cross-linked agarose beads. The innovative,...

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  • TagFP Selector

    TagFP Selector

    TagFP Selector is based on a high-affinity single-domain antibody (sdAb) that is covalently immobilized on 4% cross-linked agarose beads. The innovative,...

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  • FluoTag®-X4 anti-GFP

    FluoTag®-X4 anti-GFP

    FluoTag®-X4 anti-GFP is a blend of two in-house developed single-domain antibodies (sdAbs) that recognize two distinct epitopes on GFP...

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  • FluoTag®-X4 anti-RFP

    FluoTag®-X4 anti-RFP

    FluoTag®-X4 anti-RFP is a blend of two in-house developed single-domain antibodies (sdAbs) recognizing the most common red fluorescent proteins...

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  • FluoTag®-X2 anti-TagFP

    FluoTag®-X2 anti-TagFP

    FluoTag®-X2 anti-TagFP is derived from an in-house developed single-domain antibody (sdAb) that recognizes fluorescent proteins deriving from Entacmaea quadricolor,...

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  • FluoTag®-Q anti-GFP

    FluoTag®-Q anti-GFP

    FluoTag®-Q anti-GFP is derived from an in-house developed single-domain antibody (sdAb) that recognizes GFP and its most common derivatives...

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  • FluoTag®-Q anti-RFP

    FluoTag®-Q anti-RFP

    FluoTag®-Q anti-RFP is derived from an in-house developed single-domain antibody (sdAb) recognizing the most common red fluorescent proteins like...

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  • FluoTag®-Q anti-TagFP

    FluoTag®-Q anti-TagFP

    FluoTag®-Q anti-TagFP is derived from an in-house developed single-domain antibody (sdAb) that recognizes fluorescent proteins deriving from Entacmaea quadricolor,...

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  • MBP Selector

    MBP Selector

    MBP Selector is based on a high-affinity single-domain antibody (sdAb) that is covalently immobilized on 4% cross-linked agarose beads. The innovative,...

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  • GST Selector

    GST Selector

    GST Selector is based on a high-affinity single-domain antibody (sdAb) that is covalently immobilized on 4% cross-linked agarose beads. The innovative,...

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