OK Isolectin GS-IB but Not Very Specific

Orthopedic surgery
Johns Hopkins University School of Medicine
Postdoctoral Researcher

Overall

Quality of Results

Ease-of-Optimization

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Company:

Invitrogen

Product Name:

Isolectin GS-IB4, Alexa Fluor 488 conjugate

Catalog Number:

I21411

Clone Number:

N/A

My research focuses on molecular mechanisms of axon growth and guidance during neural development and axon regeneration after injuries to the central nervous system or peripheral nerves. For this experiment, I needed to do immunofluorescent staining on Dorsal root ganglion cell culture.

Experimental Design and Results Summary

Applications

Immunofluorescence

Sample

Mouse dorsal root ganglion cell culture

Primary Incubation

1:200, overnight at 4°C

Blocking Agent

PBS containing 10% goat serum

Secondary Incubation

N/A

Tertiary Incubation

N/A

Detection

CCD camera connected to a Zeiss inverted fluorescence microscope controlled by AxioVision software

Results Summary

IB4 just binds to the cell membrane, so you should stain IB4 individually in no-Triton incubation buffer before using any other antibodies for staining cytoplasm molecules. However, it is hard to adjust the concentration of this antibody. The signal is weak when low concentration is applied or non-specific when in high concentration. Sometimes you can get good picture but the result is not consistently good.

DOI or PMID #

N/A

Additional Notes

N/A

Image Gallery

Summary

The Good

The antibody is ok

The Bad

Not very specific

The Bottom Line

You should do some pre-experiments for getting a proper concentration.

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