ez link phosphine peg3 biotin  (Millipore)


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  • 85

    Structured Review

    Millipore ez link phosphine peg3 biotin
    Ez Link Phosphine Peg3 Biotin, supplied by Millipore, used in various techniques. Bioz Stars score: 85/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/ez link phosphine peg3 biotin/product/Millipore
    Average 85 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    ez link phosphine peg3 biotin - by Bioz Stars, 2019-12
    85/100 stars

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    Related Articles

    Sample Prep:

    Article Title: Mining the cellular inventory of pyridoxal phosphate-dependent enzymes with functionalized cofactor mimics
    Article Snippet: Paragraph title: Sample preparation and MS-based proteomic analysis ... PL3 -labeled samples were instead conjugated to EZ-Link™ Phosphine-PEG3 -Biotin (Sigma Aldrich) via Staudinger ligation (0.2 mM per 1 ml sample, 2 mg protein, 4 h at 37°C, then 20 h at 25°C).

    In Situ:

    Article Title: Mining the cellular inventory of pyridoxal phosphate-dependent enzymes with functionalized cofactor mimics
    Article Snippet: ( 2h in situ protocol ): A stationary phase culture of S. aureus USA300 TnPdxS (Nebraska transposon mutant library) grown in B-medium was exchanged into chemically-defined (PL-free) medium (CDM) containing 5 µg/ml Erythromycin and adjusted to OD600 = 40. .. PL3 -labeled samples were instead conjugated to EZ-Link™ Phosphine-PEG3 -Biotin (Sigma Aldrich) via Staudinger ligation (0.2 mM per 1 ml sample, 2 mg protein, 4 h at 37°C, then 20 h at 25°C).

    Ligation:

    Article Title: Mining the cellular inventory of pyridoxal phosphate-dependent enzymes with functionalized cofactor mimics
    Article Snippet: Following protein precipitation, samples were solubilized (PBS containing 0.4% SDS) and conjugated by CuAAC to a tri-functional rhodamine-biotin-azide. .. PL3 -labeled samples were instead conjugated to EZ-Link™ Phosphine-PEG3 -Biotin (Sigma Aldrich) via Staudinger ligation (0.2 mM per 1 ml sample, 2 mg protein, 4 h at 37°C, then 20 h at 25°C). .. Upon avidin-bead enrichment, samples were reduced with 5 mM TCEP (1 h at 37°C), alkylated using 10 mM IAA (30 min at 25°C) and quenched with 10 mM DTT (30 min at 25°C) on-bead.

    Mutagenesis:

    Article Title: Mining the cellular inventory of pyridoxal phosphate-dependent enzymes with functionalized cofactor mimics
    Article Snippet: ( 2h in situ protocol ): A stationary phase culture of S. aureus USA300 TnPdxS (Nebraska transposon mutant library) grown in B-medium was exchanged into chemically-defined (PL-free) medium (CDM) containing 5 µg/ml Erythromycin and adjusted to OD600 = 40. .. PL3 -labeled samples were instead conjugated to EZ-Link™ Phosphine-PEG3 -Biotin (Sigma Aldrich) via Staudinger ligation (0.2 mM per 1 ml sample, 2 mg protein, 4 h at 37°C, then 20 h at 25°C).

    Labeling:

    Article Title: Mining the cellular inventory of pyridoxal phosphate-dependent enzymes with functionalized cofactor mimics
    Article Snippet: Following protein precipitation, samples were solubilized (PBS containing 0.4% SDS) and conjugated by CuAAC to a tri-functional rhodamine-biotin-azide. .. PL3 -labeled samples were instead conjugated to EZ-Link™ Phosphine-PEG3 -Biotin (Sigma Aldrich) via Staudinger ligation (0.2 mM per 1 ml sample, 2 mg protein, 4 h at 37°C, then 20 h at 25°C). .. Upon avidin-bead enrichment, samples were reduced with 5 mM TCEP (1 h at 37°C), alkylated using 10 mM IAA (30 min at 25°C) and quenched with 10 mM DTT (30 min at 25°C) on-bead.

    Incubation:

    Article Title: Mining the cellular inventory of pyridoxal phosphate-dependent enzymes with functionalized cofactor mimics
    Article Snippet: For each sample, 1 ml of this suspension (≈ 1 mg protein) was incubated with 10 µl probe or DMSO (100× stock in DMSO) for 2 h at 37°C and bacteria were washed with PBS prior to cell lysis. ( ): Overnight cultures of S. aureus USA300 TnPdxS grown in B-medium were washed in CDM-Erm and used to inoculate cultures (to OD600 = 0.1) in CDM-Erm (100 ml) supplemented with 25 µM probe (25 µl of 100 mM stock in DMSO) or 0.5 µM PL (25 µl of 2 mM stock in DMSO) in baffled flasks. .. PL3 -labeled samples were instead conjugated to EZ-Link™ Phosphine-PEG3 -Biotin (Sigma Aldrich) via Staudinger ligation (0.2 mM per 1 ml sample, 2 mg protein, 4 h at 37°C, then 20 h at 25°C).

    Mass Spectrometry:

    Article Title: Mining the cellular inventory of pyridoxal phosphate-dependent enzymes with functionalized cofactor mimics
    Article Snippet: Paragraph title: Sample preparation and MS-based proteomic analysis ... PL3 -labeled samples were instead conjugated to EZ-Link™ Phosphine-PEG3 -Biotin (Sigma Aldrich) via Staudinger ligation (0.2 mM per 1 ml sample, 2 mg protein, 4 h at 37°C, then 20 h at 25°C).

    Lysis:

    Article Title: Mining the cellular inventory of pyridoxal phosphate-dependent enzymes with functionalized cofactor mimics
    Article Snippet: Bacteria were grown to stationary phase (30 h at 37°C), harvested, washed with PBS and adjusted to 1 ml OD600 = 60 (≈ 2 mg protein) prior to cell lysis. .. PL3 -labeled samples were instead conjugated to EZ-Link™ Phosphine-PEG3 -Biotin (Sigma Aldrich) via Staudinger ligation (0.2 mM per 1 ml sample, 2 mg protein, 4 h at 37°C, then 20 h at 25°C).