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netn-100 buffer  (Merck KGaA)

 
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    Structured Review

    Merck KGaA netn-100 buffer
    Netn 100 Buffer, supplied by Merck KGaA, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/netn-100+buffer/netn+100+buffer/pmc08045150-38-11-38
    Average 90 stars, based on 1 article reviews
    netn-100 buffer - by Bioz Stars, 2026-10
    90/100 stars

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    Related Articles

    Protease Inhibitor:

    Article Title: Ring finger 20/ring finger 40/WW domain-containing adaptor with coiled-coil complex interacts with p53 to regulate gene transcription in DNA damage response
    Article Snippet: After washing with 1X PBS, the cells were lysed with the NETN-100 buffer [150 mM NaCl, 0.2 mM EDTA, 1% NP-40 and 50 mM Tris-HCl (pH 7.5)] for 10 min on ice, supplemented with 1X protease inhibitor cocktail (Merck KGaA) and 1 mM phenylmethylsulfonyl fluoride, and centrifuged at 12,000 × g at 4°C for 10 min. For Co-IP, the cell lysates were incubated with 50 μl protein A/G agarose (Sangon Biotech Co., Ltd.) and the following antibodies (dilution, 1:1,000 for western blotting and 1:500 for Co-IP): Anti-HA (cat. no. D199961-0100; Sangon Biotech Co., Ltd.), anti-p53 (cat. no. ab26; Abcam), anti-FLAG (cat. no. 14793; Cell Signaling Technology, Inc.), anti-WAC (cat. no. ABE471; Merck KGaA), anti-rat IgG (cat. no. ab172730; Abcam), anti-RNF20 (cat. no. A300-714A; Bethyl Laboratories, Inc.) and anti-RNF40 (cat. no. Q2680124; Bethyl Laboratories, Inc.) overnight at 4°C.

    Incubation:

    Article Title: Ring finger 20/ring finger 40/WW domain-containing adaptor with coiled-coil complex interacts with p53 to regulate gene transcription in DNA damage response
    Article Snippet: After washing with 1X PBS, the cells were lysed with the NETN-100 buffer [150 mM NaCl, 0.2 mM EDTA, 1% NP-40 and 50 mM Tris-HCl (pH 7.5)] for 10 min on ice, supplemented with 1X protease inhibitor cocktail (Merck KGaA) and 1 mM phenylmethylsulfonyl fluoride, and centrifuged at 12,000 × g at 4°C for 10 min. For Co-IP, the cell lysates were incubated with 50 μl protein A/G agarose (Sangon Biotech Co., Ltd.) and the following antibodies (dilution, 1:1,000 for western blotting and 1:500 for Co-IP): Anti-HA (cat. no. D199961-0100; Sangon Biotech Co., Ltd.), anti-p53 (cat. no. ab26; Abcam), anti-FLAG (cat. no. 14793; Cell Signaling Technology, Inc.), anti-WAC (cat. no. ABE471; Merck KGaA), anti-rat IgG (cat. no. ab172730; Abcam), anti-RNF20 (cat. no. A300-714A; Bethyl Laboratories, Inc.) and anti-RNF40 (cat. no. Q2680124; Bethyl Laboratories, Inc.) overnight at 4°C.

    Western Blot:

    Article Title: Ring finger 20/ring finger 40/WW domain-containing adaptor with coiled-coil complex interacts with p53 to regulate gene transcription in DNA damage response
    Article Snippet: After washing with 1X PBS, the cells were lysed with the NETN-100 buffer [150 mM NaCl, 0.2 mM EDTA, 1% NP-40 and 50 mM Tris-HCl (pH 7.5)] for 10 min on ice, supplemented with 1X protease inhibitor cocktail (Merck KGaA) and 1 mM phenylmethylsulfonyl fluoride, and centrifuged at 12,000 × g at 4°C for 10 min. For Co-IP, the cell lysates were incubated with 50 μl protein A/G agarose (Sangon Biotech Co., Ltd.) and the following antibodies (dilution, 1:1,000 for western blotting and 1:500 for Co-IP): Anti-HA (cat. no. D199961-0100; Sangon Biotech Co., Ltd.), anti-p53 (cat. no. ab26; Abcam), anti-FLAG (cat. no. 14793; Cell Signaling Technology, Inc.), anti-WAC (cat. no. ABE471; Merck KGaA), anti-rat IgG (cat. no. ab172730; Abcam), anti-RNF20 (cat. no. A300-714A; Bethyl Laboratories, Inc.) and anti-RNF40 (cat. no. Q2680124; Bethyl Laboratories, Inc.) overnight at 4°C.



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