Anti-Rabbit HRP-linked Secondary Antibody

October 03, 2017

Cancer Biology
University of South Alabama
Graduate Student

Overall

Quality of Results

Ease-of-Optimization

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Company:

Cell Signaling Technology

Product Name:

Anti-Rabbit IgG, HRP-linked Antibody

Catalog Number:

7074

This HRP-linked secondary antibody is used for the detection of proteins that are probed with a rabbit antibody. I use this secondary antibody often for the detection of GAPDH in MIA PaCa-2 human pancreatic tumor cells. The horseradish peroxidase signal is easy to detect and is specific for rabbit antibodies.

Experimental Design and Results Summary

Applications

Western Blot

Sample

MIA PaCa-2 Human Pancreatic Tumor Cell Lysate

Primary Incubation

Cell Signaling GAPDH, 1:5000 dilution, 8 hours at 4 degrees Celsius in TBS-T

Blocking Agent

5% Milk in TBS-T

Secondary Incubation

Cell Signaling Anti-Rabbit, HRP-linked secondary, 1:5000 dilution, 1 hour at room temperature in TBS-t

Tertiary Incubation

None

Detection

ThermoFisher SuperSignal West Pico ECL Substrate, BioRad ChemiDoc Imager

Results Summary

This secondary antibody gives a very intense signal for GAPDH in MIA PaCa-2 human pancreatic tumor cells. It's very easy to use and provides dark bands that are specific to rabbit primary antibodies.

DOI or PMID #

N/A

Additional Notes

N/A

Image Gallery

Summary

The Good

Very specific, intense signal, very little is used for each blot since it can be diluted 1:5000

The Bad

The signal is sometimes so intense that is can make it difficult to detect other proteins if you are probing for multiple proteins on the same membrane.

The Bottom Line

This secondary antibody is very reliable, specific, and sensitive to ECL substrates. It is very economical and effective at consistently detecting proteins that are probed with rabbit primary antibodies.

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