Products

Blocking One‐P

Features

Product Image

  • Safe - does not contain phosphorylated proteins
  • Confirmed - endogeneous phosphatase free
  • High efficiency - more effective than 1% BSA

Product Information

In immunological detection methods, blocking agents containing phosphoproteins such as skim milk are not well suited for immunoassays with phospho-specific antibodies, and cause high background. Blocking One-P is an exclusive blocking solution, free of phosphate group and endogenous phosphatase. For phospho protein detection. The performance is superior compared with conventional blocking solutions such as 1% BSA. The preservative does not affect the enzyme activity of peroxidase (POD) or alkaline phosphatase (ALP). Only simple refrigerator storage is necessary, even after opening the bottle.

Comparison

  Composition Phospho-specific
antibody applications
Blocking efficacy
Blocking One-P - High molecular weight compounds
- BSA
+++ +++
Blocking One - High molecular weight compounds
- BSA
- Casein
+ +++
Skim milk - Casein - +
1% BSA - BSA ++ +

Applications

Western Blotting with phospho-specific antibody

Sample HeLa whole cell lysate (1.0 x 107)
+ RIPA Buffef (Product No.: 08714-04) (1 ml)
Lane 1. 5 µl (SDS-PAGE Gel)
Lane 2. 10 µl (SDS-PAGE Gel)
Lane 3. 5 µl (Blotting Membrane)
Lane 4. 10 µl (Blotting Membrane)
Lane 5. 5 µl
Lane 6. 10 µl
Blocking Blocking One-P, 20 min.
Primary antibody Anti-p-Tyr(PY20)(Mouse), Monoclonal (Santa Cruz: #sc-508), 1:2000 dilution
Secondary antibody Anti-Mouse IgG-HRP (Santa Cruz: #sc-2005), 1:10000 dilution
Detection CBB Stain One (Product No.: 04543-51)
Chemi-Lumi One L (Product No.: 07880-70)

Protocols

Blocking One-P pdf(PDF 86 KB)

Downloads

Blocking One Seriespdf (PDF 1.42 MB)

Ordering Information

Product Name Storage Product No. PKG Size Price
Blocking One-P 0-10°C 05999-84 200 ml e-Nacalai
Product Name Storage Product No. PKG Size Price
Streptavidin Biotin Complex Peroxidase Kit 0-10°C 30462-30 1 Kit e-Nacalai
Chemi-Lumi One Super 0-10°C 02230-30 1 Kit
Chemi-Lumi One L 0-10°C 07880-70 1 Kit
WB Stripping Solution 0-10°C 05364-55 500 ml
WB Stripping Solution Strong 0-10°C 05677-65 500 ml
Peroxidase Stain DAB Kit (Brown Stain) 0-10°C 25985-50 1 Kit
Peroxidase Stain DAB Kit (Dark-gray Stain) 0-10°C 25986-40 1 Kit
Peroxidase Stain Kit for Immuno-blotting, Nuclease tested -20°C 26652-70 1 Kit
BCIP-NBT Solution Kit for Alkaline Phosphatase Stain, Nuclease tested -20°C 03937-60 1 Kit
ELISA POD Substrate A. B. T. S. Kit 0-10°C 14351-80 1 Kit
ELISA POD Substrate TMB Kit (Popular) 0-10°C
(Do not Freze)
05298-80 1 Kit
ELISA POD Substrate TMB Solution (Easy) 0-10°C
(Do not Freze)
05299-54 100 ml

(Storage) RT: Room temperature, A: Cool and dark, R: Refrigerator, F: Freeze