抗体信号增强剂

Overview

• 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

• 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.

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• 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. 

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• 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).
  

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• 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.