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Gst fusion protein

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The GST-fusion protein is a lab equipment used for protein purification and expression. It facilitates the attachment of a glutathione S-transferase (GST) tag to a target protein, allowing for efficient isolation and purification of the protein of interest.

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Lab products found in correlation

Glutathione agarose, by Thermo Fisher Scientific (2 mentions)

2 protocols using gst fusion protein

1

Pulldown Assay for OCRL Interaction

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Pulldown experiments with purified GST or GST-SdhA truncations were performed with the lysates of HEK cells transfected with GFP or GFP-OCRL after 24 hr transfection. The lysates were prepared from a 10 cm dish in 1ml of pull down/lysis buffer (25 mM HEPES–KOH (pH 7.2), 125 mM potassium acetate, 2.5 mM magnesium acetate, 0.4% Triton X-100, and protease inhibitor cocktail), followed by incubation for 3 hr at 4°C with 250 μg of GST-fusion protein coupled to glutathione-agarose (Thermo Scientific). Beads were then washed four times (pull down buffer containing 0.1% Triton X-100) and resuspended with SDS-PAGE sample buffer followed by SDS-PAGE and western blotting. For MS analysis, proteins were eluted with SDS sample buffer followed by fractionation on SDS-PAGE, then excised from the gel, and analyzed by LC/MS/MS by the Taplin Mass Spectrometry Facility (Harvard Medical School).
Choi W.Y., Kim S., Aurass P., Huo W., Creasey E.A., Edwards M., Lowe M, & Isberg R.R. (2021). SdhA blocks disruption of the Legionella-containing vacuole by hijacking the OCRL phosphatase. Cell reports, 37(5), 109894.
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2

Pulldown Assay for OCRL Interaction

Check if the same lab product or an alternative is used in the 5 most similar protocols
Pulldown experiments with purified GST or GST-SdhA truncations were performed with the lysates of HEK cells transfected with GFP or GFP-OCRL after 24 hr transfection. The lysates were prepared from a 10 cm dish in 1ml of pull down/lysis buffer (25 mM HEPES–KOH (pH 7.2), 125 mM potassium acetate, 2.5 mM magnesium acetate, 0.4% Triton X-100, and protease inhibitor cocktail), followed by incubation for 3 hr at 4°C with 250 μg of GST-fusion protein coupled to glutathione-agarose (Thermo Scientific). Beads were then washed four times (pull down buffer containing 0.1% Triton X-100) and resuspended with SDS-PAGE sample buffer followed by SDS-PAGE and western blotting. For MS analysis, proteins were eluted with SDS sample buffer followed by fractionation on SDS-PAGE, then excised from the gel, and analyzed by LC/MS/MS by the Taplin Mass Spectrometry Facility (Harvard Medical School).
Choi W.Y., Kim S., Aurass P., Huo W., Creasey E.A., Edwards M., Lowe M, & Isberg R.R. (2021). SdhA blocks disruption of the Legionella-containing vacuole by hijacking the OCRL phosphatase. Cell reports, 37(5), 109894.
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