Transfected HEK293 cell lysate was incubated with 500 μg of peptide F (Proteintech) or control peptide (Proteintech) overnight at 4°C. The sample was cross-linked with 2 mM DTSSP (Thermo Scientific) for 1 hour at room temperature. The sample was precleared by adding 10 μg of mouse immunoglobulin G and 20 μl of protein A/G agarose and incubated at 4°C for 2 hours. After centrifugation at 1000g for 5 min, the supernatant was incubated with 5 μg of anti-DDK antibodies (OriGene) overnight at 4°C. Protein A/G PLUS-Agarose beads (40 μl) (Santa Cruz Biotechnology) were added and incubated for 2 hours at 4°C, then centrifuged for 5 min at 1000g at 4°C. The pellet was washed three times with IP lysis buffer (Thermo Scientific), followed by addition of 80 μl of 2× sample buffer (Santa Cruz Biotechnology). The sample was boiled for 5 min and then centrifuged for 5 min at 1000g at 4°C. The supernatant was loaded onto a 15% SDS-PAGE gel and probed with antibodies to peptide F (nonreducing condition), DDK, and CD59 (both in reducing condition with 5% β-mercaptoethanol).
Peptide f
Manufactured by Proteintech
Peptide F is a lab equipment product offered by Proteintech. It is designed for the synthesis and purification of peptides. The core function of Peptide F is to facilitate the chemical assembly of amino acid sequences into peptide molecules.
Automatically generated - may contain errors
Lab products found in correlation
Ip lysis buffer, by Thermo Fisher Scientific (2 mentions)
Anti ddk antibody, by OriGene (2 mentions)
Protein a g plus agarose bead, by Santa Cruz Biotechnology (2 mentions)
Dtssp, by Thermo Fisher Scientific (2 mentions)
Control peptide, by Proteintech (2 mentions)
Sample buffer, by Santa Cruz Biotechnology (2 mentions)
2 protocols using peptide f
1
Peptide F-Protein Interaction Assay
2
Peptide F-Protein Interaction Assay
Check if the same lab product or an alternative is used in the 5 most similar protocols
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Transfected HEK293 cell lysate was incubated with 500 μg of peptide F (Proteintech) or control peptide (Proteintech) overnight at 4°C. The sample was cross-linked with 2 mM DTSSP (Thermo Scientific) for 1 hour at room temperature. The sample was precleared by adding 10 μg of mouse immunoglobulin G and 20 μl of protein A/G agarose and incubated at 4°C for 2 hours. After centrifugation at 1000g for 5 min, the supernatant was incubated with 5 μg of anti-DDK antibodies (OriGene) overnight at 4°C. Protein A/G PLUS-Agarose beads (40 μl) (Santa Cruz Biotechnology) were added and incubated for 2 hours at 4°C, then centrifuged for 5 min at 1000g at 4°C. The pellet was washed three times with IP lysis buffer (Thermo Scientific), followed by addition of 80 μl of 2× sample buffer (Santa Cruz Biotechnology). The sample was boiled for 5 min and then centrifuged for 5 min at 1000g at 4°C. The supernatant was loaded onto a 15% SDS-PAGE gel and probed with antibodies to peptide F (nonreducing condition), DDK, and CD59 (both in reducing condition with 5% β-mercaptoethanol).
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