Good Antibody for Human Beta 1 Integrin Detection

August 13, 2018

BlueskyReddit
The Cell and Molecular Biology
The Scripps Research Institute
Assistant Professor

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Company:

Novus Biologicals

Product Name:

Integrin beta 1/CD29 Antibody (MEM-101A)

Catalog Number:

NB500-383

Integrins are cell surface transmembrane molecules that primarily mediate adhesion of cells to matrix proteins and induce cell signaling. Among integrins, beta 1 integrins play crucial roles in cell communications and migration of normal and cancer cells. Beta 1 integrins are also involved in stromal invasion and distant metastasis of cancer cells. We analyzed the levels of beta 1 integrin expression in human prostate carcinoma cells exhibiting different dissemination potentials. To achieve this goal, we compared several anti-integrin beta 1 antibodies, including MEM-101A clone, for their ability to recognize beta 1 integrin in western blotting under reducing and non-reducing conditions.

Experimental Design and Results Summary

Applications

Western Blot

Sample

Lysates of human tumor cells; 20 microgram per lane

Primary Incubation

Antibody at a final concentration of 1 microgram/ml, ON incubation with gentle rocking in a cold room

Blocking Agent

5% BSA in PBS-0.05% Tween; 45-60 min, RT, gentle rocking

Secondary Incubation

HRP-conjugated anti-mouse antibody at 1:5,000 in 5% milk in PBS-0.05% Tween 20

Tertiary Incubation

N/A

Detection

West Pico

Results Summary

The mAb MEM-101A from Novus specifically detected one protein band with an apparent mol. wt. of ~130 kDa, which is expected for human beta 1 integrin. The high specificity of this antibody was confirmed by the lack of any positive signal from the cells in which beta 1 integrin was stably knocked down with beta 1-specific shRNA (beta 1-KD) compared with the signal intensity in control cells (Ctrl). The MEM-101A antibody appears to detect beta 1 integrin only under non-reducing conditions as no positive band was detected in control cells under reducing conditions (Red).

DOI or PMID #

N/A

Additional Notes

The image indicates that under non-reducing conditions, this antibody specifically recognized beta 1 integrin in control cells (Ctrl), but not in the cells with beta 1 integrin knocked down (beta1-KD).

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Summary

The Good

High specificity and sensitivity.

The Bad

The antibody appears not capable of recognizing beta 1 integrin under reducing conditions.

The Bottom Line

The antibody MEM-101A is a sensitive mouse monoclonal antibody that specifically recognizes human beta 1 integrin in western blotting under non-reducing conditions.

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