Whole cell lysates were obtained from MDS-L cell line or BM-MNCs of MDS patients, and separate the same amount of protein by 12% sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE), and then blotted onto a polyvinylidene difluoride (PVDF) membrane. The PVDF membranes were blocked with Tris-buffered saline (TBS) containing 5% skimmed milk powder for 1 h, and then incubated with the primary antibodies (Supplementary Table 2 ) at 4°C overnight. Membranes were incubated with either anti-mouse or anti-rabbit immunoglobulin G (IgG) horseradish peroxidase-conjugated secondary antibody (Amersham Biosciences, Piscataway, NJ, United States). Specific bands were visualized using ECL Western Blotting Detection Reagents (Amersham Biosciences, Piscataway, NJ, United States). The intensity of bands was quantified using Image Lab software version 2.0 (Bio-Rad Laboratories, Hercules, CA, United States), and glyceraldehyde 3-phosphate dehydrogenase (GAPDH) was used as an internal standard (Xu et al., 2014 (link)).
Igg horseradish peroxidase conjugated secondary antibody
Manufactured by Cytiva
Sourced in United States
The IgG) horseradish peroxidase-conjugated secondary antibody is a laboratory reagent used in various immunoassay techniques. It consists of a secondary antibody that has been conjugated with the enzyme horseradish peroxidase. This conjugate can be used to detect and quantify target proteins or other biomolecules in samples.
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2 protocols using igg horseradish peroxidase conjugated secondary antibody
1
Western Blot Analysis of MDS Proteins
2
Western Blot Analysis of MDS Cell Samples
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Whole cell lysates were obtained from MDS-L cell line or BM-MNCs of MDS patients, and equal quantities of protein were separated by 12% sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and blotted onto a polyvinylidene di uoride (PVDF) membrane. The PVDF membranes were blocked with Tris-buffered saline (TBS) containing 5% skimmed milk powder for 1 h, incubated with primary antibodies (Supplementary Table 2) overnight at 4℃. Membranes were incubated with either anti-mouse or antirabbit immunoglobulin G (IgG) horseradish peroxidase-conjugated secondary antibody (Amersham Biosciences, Piscataway, NJ, USA). Speci c bands were visualized using ECL Western Blotting Detection Reagents (Amersham Biosciences, Piscataway, NJ, USA). The intensity of bands was quanti ed using Image Lab software version 2.0 (Bio-Rad Laboratories, Hercules, CA, USA), and glyceraldehyde 3phosphate dehydrogenase (GAPDH) was used as an internal standard.
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