CD147 Monoclonal Antibody for ICC

Center for Hearing & Deafness
University at Buffalo
Principal Investigator

Overall

Quality of Results

Ease-of-Optimization

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Company:

Pharmingen

Product Name:

Purified Mouse Anti-Human CD147

Catalog Number:

555961

CD147 is a marker that is associated with invasion and metastasis in a variety of malignant diseases. For our study, we needed to optimize CD147 immunostaining on Y79 human retinoblastoma cells. We chose this mouse monoclonal antibody, since it would pair well with a rabbit primary antibody for double immunostaining.

Experimental Design and Results Summary

Applications

Immunofluorescence

Sample

Cytospins of Y79 human retinoblastoma cells

Primary Incubation

5.0 ug/ml in PBS with 0.05% Tween20, Room Temperature for one hour

Blocking Agent

No blocking agent

Secondary Incubation

Vector Dylight 549 anti-mouse antibody (D12549) used at 5.0 ug/ml, Room Temperature for 45 minutes

Tertiary Incubation

Vectashield mounting medium containing DAPI counterstain

Detection

Fluorescence microscopy- Nikon microscope with SPOT camera

Results Summary

In the attached image, you can see very clear fluorescent staining of CD147 on the surface of the Y79 human retinoblastoma cells (red) in conjunction with the DAPI counterstain (blue). The level of background is very low.

Additional Notes

None

Related Categories

Image Gallery

Summary

The Good

Very clear staining, easy to optimize

The Bad

None

The Bottom Line

Overall, this antibody was very easy to optimize, with the best results at 5 ug/ml. I recommend this antibody for use in fluorescent immunocytochemistry.

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