HOW DID OUR ANTIBODY PERFORM?

ANTIBODY CUSTOMER REVIEW: Anti-c-Myc Polyclonal Antibody (STJ92356)

What was used?Primary antibody: STJ92356 Anti-c-Myc Antibody

Secondary antibody: 159Tb-IgG goat anti-rabbit

Provider: St John's Laboratory

Dilution ratio: Used at 4 µg/ml

Application: Mass Cytometry

Materials for validation: MCF7, SKBR3, MDA-MB-231, T47D cell lines (ATCC)Click for product data sheet PDF

What Was The Protocol?

Treatment of Materials

The cells were cultured to 70%-80% plating density. For cell detachment, they were washed with pre-warmed PBS (Gibco) and 1X TrypLETM (Thermo Fisher) was added for 5 min at 37°C. For cell fixation, 1.6% paraformaldehyde (Thermo Fisher) was added for 10 minutes at room temperature. Then, the cells were pipetted off the plate and collected in cold full growth medium to quench the paraformaldehyde. The cells were spun down and aliquoted in PBS (Gibco) prior to storage at -80°C.

Protocol

Fixed cells were washed once with 1x PBS / 0.05% bovine serum albumin / 2mM EDTA cell staining buffer (CSB) prior to staining with the antibody (STJ92356, 4 µg/ml) in CSB for 45 min at 22°C. The cells were washed three times with CSB. Then, the secondary antibody, a goat anti-rabbit 159Tb-IgG (0.25 µg/ml), was applied in CSB for 45 min at 22°C. The cells were washed with CSB and ddH20 prior to mass cytometry acquisition.

What were the results?

Figure: Overlay histogram showing different mammary epithelial cell lines stained with STJ92356 and anti-rabbit 159Tb-IgG (orange line) or anti-rabbit 159Tb-IgG alone (blue line).

MCF7 T47DMDA231SKBR3

Coun

ts

Anti-C-MYC antibody (STJ92356)

What did the customer think?

Antibody Rating:

Testing results were provided independently by J. Wagner at the University of Zurich. They stated “At 4 µg/ml the antibody gives good signal intensities at good signal-to-noise ratios for the different cell lines used. The antibody can be used for mass cytometry analysis of C-MYC expression. The specificity of the antibody to C-MYC should be shown in an additional experiment.”

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