To blot Pds1–18myc, aliquots of cultures were removed during the G1 or Cdc20 release assays and placed on ice. Whole-cell protein extracts were prepared by NaOH treatment followed by lysis in Laemmli buffer heated to 95°C for 10 min. Clarified samples were electrophoresed on a 6% Tris-glycine gel and blotted using a semi-dry apparatus. Blots were developed with anti-myc – clone 9E10 from mouse, anti-beta actin – clone ab8224 from mouse (Abcam), and anti-mouse IgG HRP conjugated from sheep (GE Healthcare) using ECL detection (RPN3243; GE Healthcare).
Anti mouse igg hrp conjugated from sheep
Manufactured by GE Healthcare
The Anti-mouse IgG HRP conjugated from sheep is a laboratory reagent used to detect the presence of mouse immunoglobulin G (IgG) in various experimental and analytical procedures. It is a conjugate of sheep-derived anti-mouse IgG antibodies and the enzyme horseradish peroxidase (HRP). The HRP enzyme allows for colorimetric or chemiluminescent detection of the target mouse IgG molecules.
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2 protocols using anti mouse igg hrp conjugated from sheep
1
Western Blot Analysis of Pds1–18myc
2
Western Blot Analysis of Pds1–18myc
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To blot Pds1–18myc, aliquots of cultures were removed during the G1 or Cdc20 release assays and placed on ice. Whole-cell protein extracts were prepared by NaOH treatment followed by lysis in Laemmli buffer heated to 95°C for 10 min. Clarified samples were electrophoresed on a 6% Tris-glycine gel and blotted using a semi-dry apparatus. Blots were developed with anti-myc – clone 9E10 from mouse, anti-beta actin – clone ab8224 from mouse (Abcam), and anti-mouse IgG HRP conjugated from sheep (GE Healthcare) using ECL detection (RPN3243; GE Healthcare).
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