Intensify Western blot signals up to 5 times compared to traditional blotting procedures.
Protein-free solution eliminates cross-reaction with antibodies.
Reduces antigen and reagent use without the need for additional incubation steps.
pH @25C----7.2 - 7.6
Protease (P/F)
Antibody Signal Enhancer is a Western blotting antibody diluent that increases intensity of protein detection up to 5-fold compared to conventional blotting. The enhancer is a ready-to-use solution that replaces TBS-T with non-fat dry milk, or other diluent, for antibody incubation steps. It is formulated for use with HRP- and biotin-conjugated antibodies and works with both PVDF and nitrocellulose blots. Antibody Signal Enhancer is stable at room temperature and is a non-hazardous formulation. 200 ml size provides sufficient material for 10 blots. 500 ml size provides sufficient material for 25 blots. 1 L size provides sufficient material for 50 blots.
Antibody Signal Enhancer improves western blot detection. Data courtesy of Eckhard Jankowsky, Ph.D. Case Western Reserve University, Cleveland, OH. Protein samples were separated on an SDS-PAGE and transferred to PVDF paper. The PVDF paper was blocked with 8% BSA and probed with primary and secondary antibodies also in 8% BSA. The blot was developed and the image was captured. After stripping the blot, the PVDF paper was blocked with 10% milk and probed with primary and secondary antibodies in Antibody Signal Enhancer (M336). The blot was developed and the image was captured.
Comparison of Western blots with Antibody Signal Enhancer as antibody diluent versus conventional diluent. Cytoplasmic proteins (5 μg/lane) isolated from K562 cells using AMRESCO’s Cytoplasmic & Nuclear Protein Enrichment Kit (M330) were resolved on a 10% Fluorescent SPRINT NEXT GEL® (M317) (A, C, E). The total protein was viewed on a transilluminator prior to semi-dry transfer to PVDF using Rapid Transfer Buffer (N789). The blot was blocked in RapidBlock™ and then divided for overnight incubation in various antibodies diluted in TBST/5% non-fat dry milk (B1, D1, F1) or Antibody Signal Enhancer (M336) (B2, D2, F2). The blots were washed and then incubated for 1 hour in goat anti-rabbit HRP antibody diluted in the same diluent as the primary antibody. The blots were washed and detected with VisiGlo Plus™ HRP Chemiluminescent Substrate Kit (N219). Antibodies: 1:2,000 β-actin (B), 1:250 Hsp90 (D), 1:2,000 NF-кβ (F).
Antibody Signal Enhancer decreases the amount of antibody required for immunohistochemical staining while maintaining high antibody signal. MEFs (mouse embryonic fibroblasts) transfected with HA-GAPDH plasmid were grown on coverslips and fixed with 4% paraformaldehyde for 10 minutes at room temperature and permeablized with 0.2% Triton-X 100-PBS for 10 minutes at 370C. Cells were blocked in PBS-2% BSA for 1 hour at room temperature and incubated overnight in purified mouse monoclonal HA.11 antibody (Covance 16B12) diluted 1:500 in blocking solution (Control) or 1:1,000 in Antibody Signal Enhancer (AMRESCO M336). Cells were washed three times in PBS-0.2% Triton® X-100, then incubated 1 hour at room temperature in 100 μg/mL Hoechst 33342 (Life Technologies) and Alexa Fluor® Goat Anti-Mouse IgG (Life Technologies) diluted 1:1,000 in blocking buffer (Control) or Antibody Signal Enhancer (AMRESCO). Cells were washed, then mounted with SlowFade® Gold reagent (Life Technologies) before imaging with a Leica SP2 confocal microscope using an argon laser (488 nm) and two-photon excitation (760 nm). Courtesy of Mithu Majumder, Ph.D., M.B.A., Case Western Reserve University, Cleveland, Ohio.
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上海康励仪器设备有限公司
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310118002832441
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2013-04-10
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100
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