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Western blot (A) and silver-stained gel (B) show the purification fractions of the WT control (WT/WT), bait/WT experiment (bait/WT), and bait control (bait/bait), all crosslinked with 0.2% (w/v) FA. The fractions loaded are lysate (L) (1 μL), flowthrough (FT) (1 μL), wash steps 1 (W1) and 3 (W3) (8 μL), eluate (E) (4 μL), and residue (R) (4 μL). The Chameleon ® Due Pre-Stained protein marker was used for molecular weight reference. The position of the signals for PycA (127.72 kDa) and SigA-TS (45.9 kDa) is marked on the right side. PycA is a biotinylated protein and thus is purified unspecifically by the <t>MagStrep</t> beads.
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Western blot (A) and silver-stained gel (B) show the purification fractions of the WT control (WT/WT), bait/WT experiment (bait/WT), and bait control (bait/bait), all crosslinked with 0.2% (w/v) FA. The fractions loaded are lysate (L) (1 μL), flowthrough (FT) (1 μL), wash steps 1 (W1) and 3 (W3) (8 μL), eluate (E) (4 μL), and residue (R) (4 μL). The Chameleon ® Due Pre-Stained protein marker was used for molecular weight reference. The position of the signals for PycA (127.72 kDa) and SigA-TS (45.9 kDa) is marked on the right side. PycA is a biotinylated protein and thus is purified unspecifically by the <t>MagStrep</t> beads.
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Western blot (A) and silver-stained gel (B) show the purification fractions of the WT control (WT/WT), bait/WT experiment (bait/WT), and bait control (bait/bait), all crosslinked with 0.2% (w/v) FA. The fractions loaded are lysate (L) (1 μL), flowthrough (FT) (1 μL), wash steps 1 (W1) and 3 (W3) (8 μL), eluate (E) (4 μL), and residue (R) (4 μL). The Chameleon ® Due Pre-Stained protein marker was used for molecular weight reference. The position of the signals for PycA (127.72 kDa) and SigA-TS (45.9 kDa) is marked on the right side. PycA is a biotinylated protein and thus is purified unspecifically by the <t>MagStrep</t> beads.
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magstrep streptactin xt beads  (IBA Lifesciences)
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Western blot (A) and silver-stained gel (B) show the purification fractions of the WT control (WT/WT), bait/WT experiment (bait/WT), and bait control (bait/bait), all crosslinked with 0.2% (w/v) FA. The fractions loaded are lysate (L) (1 μL), flowthrough (FT) (1 μL), wash steps 1 (W1) and 3 (W3) (8 μL), eluate (E) (4 μL), and residue (R) (4 μL). The Chameleon ® Due Pre-Stained protein marker was used for molecular weight reference. The position of the signals for PycA (127.72 kDa) and SigA-TS (45.9 kDa) is marked on the right side. PycA is a biotinylated protein and thus is purified unspecifically by the <t>MagStrep</t> beads.
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Western blot (A) and silver-stained gel (B) show the purification fractions of the WT control (WT/WT), bait/WT experiment (bait/WT), and bait control (bait/bait), all crosslinked with 0.2% (w/v) FA. The fractions loaded are lysate (L) (1 μL), flowthrough (FT) (1 μL), wash steps 1 (W1) and 3 (W3) (8 μL), eluate (E) (4 μL), and residue (R) (4 μL). The Chameleon ® Due Pre-Stained protein marker was used for molecular weight reference. The position of the signals for PycA (127.72 kDa) and SigA-TS (45.9 kDa) is marked on the right side. PycA is a biotinylated protein and thus is purified unspecifically by the <t>MagStrep</t> beads.
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Western blot (A) and silver-stained gel (B) show the purification fractions of the WT control (WT/WT), bait/WT experiment (bait/WT), and bait control (bait/bait), all crosslinked with 0.2% (w/v) FA. The fractions loaded are lysate (L) (1 μL), flowthrough (FT) (1 μL), wash steps 1 (W1) and 3 (W3) (8 μL), eluate (E) (4 μL), and residue (R) (4 μL). The Chameleon ® Due Pre-Stained protein marker was used for molecular weight reference. The position of the signals for PycA (127.72 kDa) and SigA-TS (45.9 kDa) is marked on the right side. PycA is a biotinylated protein and thus is purified unspecifically by the <t>MagStrep</t> beads.
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Western blot (A) and silver-stained gel (B) show the purification fractions of the WT control (WT/WT), bait/WT experiment (bait/WT), and bait control (bait/bait), all crosslinked with 0.2% (w/v) FA. The fractions loaded are lysate (L) (1 μL), flowthrough (FT) (1 μL), wash steps 1 (W1) and 3 (W3) (8 μL), eluate (E) (4 μL), and residue (R) (4 μL). The Chameleon ® Due Pre-Stained protein marker was used for molecular weight reference. The position of the signals for PycA (127.72 kDa) and SigA-TS (45.9 kDa) is marked on the right side. PycA is a biotinylated protein and thus is purified unspecifically by the MagStrep beads.

Journal: Bio-protocol

Article Title: TIE-UP-SIN: A Method for Enhanced Identification of Protein–Protein Interactions

doi: 10.21769/BioProtoc.5663

Figure Lengend Snippet: Western blot (A) and silver-stained gel (B) show the purification fractions of the WT control (WT/WT), bait/WT experiment (bait/WT), and bait control (bait/bait), all crosslinked with 0.2% (w/v) FA. The fractions loaded are lysate (L) (1 μL), flowthrough (FT) (1 μL), wash steps 1 (W1) and 3 (W3) (8 μL), eluate (E) (4 μL), and residue (R) (4 μL). The Chameleon ® Due Pre-Stained protein marker was used for molecular weight reference. The position of the signals for PycA (127.72 kDa) and SigA-TS (45.9 kDa) is marked on the right side. PycA is a biotinylated protein and thus is purified unspecifically by the MagStrep beads.

Article Snippet: MagStrep ® Strep-Tactin ® XT beads (IBA Lifesciences, catalog number: 2-5090-010) 14.

Techniques: Western Blot, Staining, Purification, Control, Residue, Marker, Molecular Weight