Total cell lysate from HeLa (15 µg/lane) were resolved by electrophoresis (4-20% Tris-Glycine), transferred to nitrocellulose, and probed with anti-β-actin antibody (clone Poly6221, rabbit IgG) (lane 1, 2, 3 and 5) and purified anti-β actin antibody as the negative control (clone 2F1-1, mouse IgG) (lane 4). After a 30 minutes wash, lanes 1 and 4, 2, 3 were probed respectively with 1:5000, 1:10000 and 1:20000 HRP anti-rabbit IgG antibodies (clone 6B9G9), and lane 5 was probed with 1:3000 HRP Donkey anti-rabbit IgG (minimal x-reactivity) antibody (clone Poly4064) as a positive control. Proteins were visualized using chemiluminescence detection system. Lane M contains the molecular weight ladder.
Total cell lysate from HeLa (15 µg/lane) were resolved by electrophoresis (4-20% Tris-Glycine), transferred to nitrocellulose, and probed with anti-β-actin antibody (clone Poly6221, rabbit IgG) (lane 1, 2, 3 and 5) and purified anti-β actin antibody as the negative control (clone 2F1-1, mouse IgG) (lane 4). After a 30 minutes wash, lanes 1 and 4, 2, 3 were probed respectively with 1:5000, 1:10000 and 1:20000 HRP anti-rabbit IgG antibodies (clone 6B9G9), and lane 5 was probed with 1:3000 HRP Donkey anti-rabbit IgG (minimal x-reactivity) antibody (clone Poly4064) as a positive control. Proteins were visualized using chemiluminescence detection system. Lane M contains the molecular weight ladder.
Optimal concentration of usage is 1:10,000 – 1:40,000 for WB. The optimal concentration should be determined by titration for each individual assay of interest.
Product Citations
Laure A, et al. 2023. Cancers (Basel). 15:. PubMed
IgG antibodies are large molecules of about 150 kD made of four peptide chains. It contains two identical class γ heavy chains of about 50 kD and two identical light chains of about 25 kD.
Distribution
Secreted, B cell surface.
Function
Antibodies are major components of humoral immunity. IgG is the main type of antibody found in blood and extracellular fluid allowing it to control infection of body tissues. By binding many kinds of pathogens such as viruses, bacteria, and fungi, IgG protects the body from infection.
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