Blocking One‐P
Features
- 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 86 KB)
Downloads
Blocking One Series (PDF 1.42 MB)
Ordering Information
| Product Name | Storage | Product No. | PKG Size | Price |
|---|---|---|---|---|
| Blocking One-P | 0-10°C | 05999-84 | 200 ml |  |
| Product Name | Storage | Product No. | PKG Size | Price |
|---|---|---|---|---|
| Streptavidin Biotin Complex Peroxidase Kit | 0-10°C | 30462-30 | 1 Kit | |
| 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
