Placental proteins were extracted as described above. After removal of the organic (lipid) phase, the protein was pelleted in methanol and dissolved in RIPA buffer overnight. After sonication of lysate, protein concentration was determined using a Pierce BCA protein assay kit (Thermo Fisher Scientific Inc. Waltham, MA). Proteins were resolved in 4–15% SDS-PAGE and transferred to polyvinylidene difluoride membranes using Trans-Blot Turbo RTA Midi PVDF Transfer Kit (Bio-Rad). Membranes were blocked in 5% BSA/TBS-T followed by overnight incubation in primary antibodies against GLUT1, LPL, CD36, FABP3, and ADRP followed by one-hour incubation in HRP-conjugated goat anti-rabbit IgG secondary antibody (SouthernBiotech, Birmingham, AL). Luminata Forte Western HRP substrate (EMD Millipore, Billerica, MA) and VisionWorks LS software (UVP, Upland, CA) were used for imaging and recording integrated optical densities. Readings were normalized to β-actin. Antibody information is shown in Supplemental Table S2 .
Hrp conjugated goat anti rabbit igg secondary antibody
Manufactured by Southern Biotech
The HRP-conjugated goat anti-rabbit IgG secondary antibody is a laboratory reagent used for the detection and quantification of rabbit primary antibodies in various immunoassays. The antibody is conjugated with the enzyme Horseradish Peroxidase (HRP), which can produce a colorimetric or chemiluminescent signal when exposed to the appropriate substrate, allowing for the visualization and measurement of the target analyte.
Automatically generated - may contain errors
2 protocols using hrp conjugated goat anti rabbit igg secondary antibody
1
Placental Protein Extraction and Analysis
2
Placental Protein Extraction and Analysis
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Placental proteins were extracted as described above. After removal of the organic (lipid) phase, the protein was pelleted in methanol and dissolved in RIPA buffer overnight. After sonication of lysate, protein concentration was determined using a Pierce BCA protein assay kit (Thermo Fisher Scientific Inc. Waltham, MA). Proteins were resolved in 4–15% SDS-PAGE and transferred to polyvinylidene difluoride membranes using Trans-Blot Turbo RTA Midi PVDF Transfer Kit (Bio-Rad). Membranes were blocked in 5% BSA/TBS-T followed by overnight incubation in primary antibodies against GLUT1, LPL, CD36, FABP3, and ADRP followed by one-hour incubation in HRP-conjugated goat anti-rabbit IgG secondary antibody (SouthernBiotech, Birmingham, AL). Luminata Forte Western HRP substrate (EMD Millipore, Billerica, MA) and VisionWorks LS software (UVP, Upland, CA) were used for imaging and recording integrated optical densities. Readings were normalized to β-actin. Antibody information is shown in Supplemental Table S2 .
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