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  3. Protein Biology
  4. Protein Purification
  5. Agarose Beads
  6. Glutathione Agarose

Glutathione Agarose 

Cat. No.: HY-K0211
Manual COA SDS Technical Support

MCE Glutathione Agarose, a 4% highly cross-linked agarose reagent, enables high-yield, high-purity purification of GST-tagged proteins.

Glutathione Agarose
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    Glutathione Agarose purchased from MedChemExpress. Usage Cited in: J Med Virol. 2022 Sep 16.  [Abstract]

    GST pull‐down assays to detect ACE2‐RBD association in vitro. First, the GST‐ACE2 and PRMT5 methylated‐GST‐ACE2 were pulled down with the HEK‐293T‐GFP‐RBD lysates, respectively. Then, the GST‐ACE2 SDMA midofication and GFP‐RBD binding with GST‐ACE2 or methylated‐GST‐ACE2 were detected by western blot, respectively. ACE2, angiotensin‐converting enzyme 2; MS, mass spectrometry; PRMT, protein arginine methyltransferases; RBD, receptor‐binding domain; SDMA, symmetric dimethyl‐arginines.

    • Description

    • Storage

    • Protocol

    • Components

    • Documentation

    Description
    & Advantages

    MCE Glutathione Agarose, a 4% highly cross-linked agarose reagent, effectively purifies high levels of overexpressed GST-tagged fusion proteins at a variety of scales. MCE Glutathione Agarose has high protein-binding capacity and stability, and is compatible with a wide range of chemicals and pH values, making it ideal for high performance purification of GST-tagged fusion proteins expressed in E. coli, yeast, insect and mammalian expression systems.

     

    The specifications of the product correspond to the actual resin volume, with the resin content of 50%.
    Storage

    2-8°C, 2 years.
    Protocol

    1. Recommended Buffers

    Binding/Wash Buffer, Elution Buffer.

    2. Load of Glutathione Agarose

    (1) Mix the slurry by gently inverting the bottle several times to completely suspend the agarose.

    (2) Transfer an appropriate-sized slurry to the column and allow the agarose to settle down and the storage buffer to drain from the column.

    (3) Equilibrate the column with 3× bed volumes of ultrapure water

    (4) Close the bottom outlet of the column

    3. Procedure

    (1) Equilibrate the column of agarose with 5× bed volumes of Binding/Wash Buffer.

    (2) Apply appropriate-sized sample to the column. If the sample contains insoluble matter, centrifuge or filter (0.22 μm or 0.45 μm filter) before use.

    Note: Binding capacity is flow rate-and protein-dependent. Higher flow rates will decrease production time, but may result in losing a small portion of the target protein.

    (3) Collect and save the flow-through for analysis.

    (4) Wash the column with approximately 10-15× bed volumes of Binding/Wash Buffer or until the absorbance is stable. If desired, save supernatant for downstream analysis.

    (5) Elute with approximately 5-10× bed volumes of Elution Buffer, collect fractions.

    (6) Analyze the target protein by SDS-PAGE, along with fractions collected from different steps if necessary

    4. Cleaning-in Place, CIP

    (1) To remove precipitated or denatured proteins, wash the column with 2× bed volumes of 6 M guanidine hydrochloride followed by 5× bed volumes of PBS (pH 7.4) washed.

    (2) To remove hydrophobic substances, wash the column with 3-4× bed volumes of 70% ethanol or 2× bed volumes of 1% Triton X-100 followed by 5× bed volumes of PBS (pH 7.4) washed.

    (3) Store agarose in 20% ethanol at 2-8ºC.
    Components
    Components HY-K0211- 5 mL HY-K0211- 10 mL HY-K0211- 50 mL
    Glutathione Agarose (Settled Resin) 5 mL 10 mL 50 mL
    Documentation

    Manual (130 KB) SDS (189 KB)
    Help & FAQs
    • Do most proteins show cross-species activity?

      Species cross-reactivity must be investigated individually for each product. Many human cytokines will produce a nice response in mouse cell lines, and many mouse proteins will show activity on human cells. Other proteins may have a lower specific activity when used in the opposite species.

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