Protocol specific for Blimp-1 Antibody (NB600-235)

Western Blot
1. Run ~20 micrograms of protein on a 10% SDS-PAGE.

2. Block nitrocellulose membrane with 5% blotto in TBS-0.2% Tween-20, overnight at 4 degrees Celcius.

3. Incubate membrane with diluted anti-Blimp-1 (primary antibody, cat# NB 600-235) for 1 hour at RT, in 2% blotto in TBS-0.2% Tween-20.

4. Wash blot 4 times for 10 minutes, each, at RT with TBS-0.2% Tween-20.

5. Incubate membrane with anti-mouse IgG1 conjugated to HRP (secondary antibody), diluted in 2% blotto in TBS-0.2% Tween-20, for 45 minutes at RT.

6. Wash 4 times for 10 minutes, each, at RT with TBS-0.2% Tween-20.

7. Incubate membrane with chemiluminescence reagent as per vendor specification. Dry blot and cover with saran wrap.

8. Expose to film.

9.Develop image.

Immunohistochemistry / Immunofluorescence
Please refer to the following refernence:
Cattoretti, G., et al. PRDM1/Blimp-1 is expressed in human B-lymphocytes committed to the plasma cell lineage.J Pathol. 206(1):76-86 (2005)