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Strep tactin superflow plus beads

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Strep-Tactin Superflow Plus beads are a type of affinity chromatography resin designed for the purification of proteins tagged with the Strep-tag II affinity tag. The beads provide a high-capacity, high-specificity binding matrix for the rapid and efficient capture and purification of Strep-tagged proteins from complex samples.

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Lab products found in correlation

Biolock, by Qiagen (2 mentions) Cytobuster protein extraction reagent, by Merck Group (2 mentions) Rnase inhibitor, by New England Biolabs (2 mentions)

Close spelling variants of this product

Strep-Tactin Superflow Plus Strep-Tactin beads Strep-Tactin

4 protocols using strep tactin superflow plus beads

1

Recombinant Ebola Virus VP40 Purification

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VP40 was expressed in Human Embryonic Kidney (HEK293T) cells using the pTriEx-5 vector (Novagen). An N-terminal Strep-tag prefix (sequence MASWSHPQFEKGADDDDK) is followed by the full-length VP40 of Zaire Ebola virus (Ebola virus/H.sapiens-tc/COD/1976/Yambuku-Mayinga, GenBank KR063671), employed as wild-type or as the mutants R134A, I307R, and ΔCTD (truncated after residue 194). A control plasmid expressed the Strep-tag prefix only. Except as stated, we co-transfected an Ebola minigenome replicon system (Jasenosky et al., 2010 (link)). Each 100 mm dish was transfected using TransIT-LT1 Transfection Reagent (Mirus), the VP40 vector (2 μg), and the vectors of the minigenome replicon system: pCEZ-NP-2A-VP35 (2.07 μg), pCEZ-VP30 (1.24 μg), pCEZ-L (12.4 μg), and pHH21–3E5E-Luc (1.04 μg). Cells were lysed after 19.5 hours using CytoBuster Protein Extraction Reagent (Millipore 71009) mixed with RNase Inhibitor (New England BioLabs M0307S). The lysate was combined with BioLock (IBA 2–0205-250), incubated 60 minutes with ~100 μL QIAGEN Strep-Tactin Superflow Plus beads using the batch method, washed twice with 50 mM sodium phosphate monobasic pH 8, 100 mM NaCl, and eluted twice for ~2 minutes using 10 mM biotin.
Landeras-Bueno S., Wasserman H., Oliveira G., VanAernum Z.L., Busch F., Salie Z.L., Wysocki V.H., Andersen K, & Saphire E.O. (2021). Cellular mRNA triggers structural transformation of Ebola virus matrix protein VP40 to its essential regulatory form. Cell reports, 35(2), 108986.
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2

Recombinant Ebola Virus VP40 Purification

Check if the same lab product or an alternative is used in the 5 most similar protocols
VP40 was expressed in Human Embryonic Kidney (HEK293T) cells using the pTriEx-5 vector (Novagen). An N-terminal Strep-tag prefix (sequence MASWSHPQFEKGADDDDK) is followed by the full-length VP40 of Zaire Ebola virus (Ebola virus/H.sapiens-tc/COD/1976/Yambuku-Mayinga, GenBank KR063671), employed as wild-type or as the mutants R134A, I307R, and ΔCTD (truncated after residue 194). A control plasmid expressed the Strep-tag prefix only. Except as stated, we co-transfected an Ebola minigenome replicon system (Jasenosky et al., 2010 (link)). Each 100 mm dish was transfected using TransIT-LT1 Transfection Reagent (Mirus), the VP40 vector (2 μg), and the vectors of the minigenome replicon system: pCEZ-NP-2A-VP35 (2.07 μg), pCEZ-VP30 (1.24 μg), pCEZ-L (12.4 μg), and pHH21–3E5E-Luc (1.04 μg). Cells were lysed after 19.5 hours using CytoBuster Protein Extraction Reagent (Millipore 71009) mixed with RNase Inhibitor (New England BioLabs M0307S). The lysate was combined with BioLock (IBA 2–0205-250), incubated 60 minutes with ~100 μL QIAGEN Strep-Tactin Superflow Plus beads using the batch method, washed twice with 50 mM sodium phosphate monobasic pH 8, 100 mM NaCl, and eluted twice for ~2 minutes using 10 mM biotin.
Landeras-Bueno S., Wasserman H., Oliveira G., VanAernum Z.L., Busch F., Salie Z.L., Wysocki V.H., Andersen K, & Saphire E.O. (2021). Cellular mRNA triggers structural transformation of Ebola virus matrix protein VP40 to its essential regulatory form. Cell reports, 35(2), 108986.
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3

Affinity Purification of Protein Complexes

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We homogenized heads, either from adult flies or from late pupae (~90% pupal stage), and used Strep-Tactin Superflow plus beads (Qiagen) to precipitate the SBP tagged bait proteins and the associated complex. We eluted the bound proteins from the resin with desthiobiotin, fractionated the proteins by SDS-PAGE and performed Western blot analyses.
Chen Z., Chen H.C, & Montell C. (2015). TRP and rhodopsin transport depends on dual XPORT ER chaperones encoded by an operon. Cell reports, 13(3), 573-584.
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4

XPORT-B Protein Purification and Interactors

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We crushed heads from wild-type (Canton-S) flies, and from xportΔB mutant flies that expressed the xport-B∷SBP transgene, and incubated the supernatants with Strep-Tactin Superflow plus beads (Qiagen). We eluted the XPORT-B∷SBP protein and its associated complex in buffer containing 1% Triton X-100 and 2.5 mM desthiobiotin, which we used for silver staining and mass spectrometry (Biomolecular and Proteomics Mass Spectrometry Facility, University of California, San Diego).
Chen Z., Chen H.C, & Montell C. (2015). TRP and rhodopsin transport depends on dual XPORT ER chaperones encoded by an operon. Cell reports, 13(3), 573-584.
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