Abcam
Anti-GAPDH Antibody
ab9485
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My research focuses on breast cancer microenvironment. In this experiment, I performed Western blotting for different proteins and used GAPDH as a reference gene and loading control, in order to verify that the amounts of cell lysate in each lane were comparable.
Western blotting
Human breast adenocarcinoma cells
Anti-GAPDH, incubated overnight (~16 hours) at 4°C. Diluted 1:10,000 in the blocking agent.
4% BSA in TBST (Tris Buffered Saline with 0.05% Tween), 1 hour at room temperature (25°C).
Peroxidase-conjugated Affinipure goat anti rabbit IgG (Cat# 111-035-003), from Jackson Immunoresearch, incubated for 1 hour in room temperature (25°C). Diluted 1:8,000 in the blocking buffer.
N/A
Self-made ECL reagents, detected with super rx-n medical x-ray film (Fuji, Cat# 47410 19284).
The antibody shows a single specific band of the expected size. Background levels are low. The results show that the amount of cell lysate was extremely similar in all lanes.
None.
Strong signal, low background, can be re-used, used in low concentrations.
The antibody is a good choice for using a reference gene since it is of good quality and is extremely cost-effective.