Anti-GAPDH Antibody from Abcam

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Cell Research
Tel Aviv University
PhD Student
Human breast adenocarcinoma whole-cell lysates were loaded and subjected to SDS-PAGE and Western blotting. The antibody was used in order to verify that the amount of cell lysate in each lane was comparable.

Company:

Abcam

Product Name:

Anti-GAPDH Antibody

Catalog Number:

ab9485

Image

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.

Experimental Design and Results Summary

Applications

Western blotting

Sample

Human breast adenocarcinoma cells

Primary Incubation

Anti-GAPDH, incubated overnight (~16 hours) at 4°C. Diluted 1:10,000 in the blocking agent.

Blocking Agent

4% BSA in TBST (Tris Buffered Saline with 0.05% Tween), 1 hour at room temperature (25°C).

Secondary Incubation

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.

Tertiary Incubation

N/A

Detection

Self-made ECL reagents, detected with super rx-n medical x-ray film (Fuji, Cat# 47410 19284).

Results Summary

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.

Additional Notes

None.

Related Categories

Image Gallery

Human breast adenocarcinoma whole-cell lysates were loaded and subjected to SDS-PAGE and Western blotting. The antibody was used in order to verify that the amount of cell lysate in each lane was comparable.

Summary

The Good

Strong signal, low background, can be re-used, used in low concentrations.

The Bad

None.

The Bottom Line

The antibody is a good choice for using a reference gene since it is of good quality and is extremely cost-effective.

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