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Our Abpromise guarantee covers the use of ab3329 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|WB||Use a concentration of 1 - 3 µg/ml. Detects a band of approximately 20 kDa.Can be blocked with Human Proteasome 20S LMP7 peptide (ab4945).|
|IHC-P||Use a concentration of 1 µg/ml.|
|ICC/IF||Use a concentration of 5 µg/ml.|
|IHC-Fr||1/500. PubMed: 17008387|
|Flow Cyt||Use 3-5µg for 106 cells.|
Detected by chemiluminescence.
Immunocytochemistry/ Immunofluorescence analysis of HeLa cells labeling Proteasome 20S LMP7 with ab3329 at 5µg/ml. The cells were fixed with 4% paraformaldehyde for 15 minutes, permeabilized with 0.1% Triton X-100 in TBS for 10 minutes, and blocked with 3% Blocker BSA in PBS for 15 minutes at room temperature. Cells were stained with or without Anti-Proteasome 20S LMP7 antibody (ab3329), at a concentration of 5µg/ml for 1 hour at room temperature, and then incubated with a Alexa Fluor® 488 goat anti-rabbit IgG secondary antibody at a dilution of 1/1000 for 1 hour s at room temperature (both panels, green). Nuclei (both panels, blue) were stained with Hoechst 33342 dye.
Flow Cytometry analysis of SH-SY5Y cells labeling Proteasome 20S LMP7 with ab3329. Cells were fixed with 70% ethanol for 10 minutes, permeabilized with 0.25% Triton™ X-100 for 20 minutes, and blocked with 5% BSA for 30 minutes at room temperature. Cells were labeled with Anti-Proteasome 20S LMP7 antibody (ab3329) (red histogram) or with rabbit isotype control (pink histogram) at 3-5 ug/million cells in 2.5% BSA. After incubation at room temperature for 2 hours, the cells were labeled with Alexa Fluor® 488 Goat Anti-Rabbit Secondary Antibody at a dilution of 1/400 for 30 minutes at room temperature. The representative 10,000 cells were acquired and analyzed for each sample. The purple histogram represents unstained control cells and the green histogram represents no-primary-antibody control.