Blocking / Detection of Phosphorylated Proteins
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.
Safe: does not contain phosphorylated proteins
Confirmed: endogenous phosphatase free
High efficiency: more effective than 1% BSA
Sample | : | HeLa whole cell lysate (1.0 x 107) + RIPA Buffer (Cat No.: 08714-04) (1 ml) Lane 1. 5 ul (SDS-PAGE Gel) Lane 2. 10 ul (SDS-PAGE Gel) Lane 3. 5 ul (Blotting Membrane) Lane 4. 10 ul (Blotting Membrane) Lane 5. 5 ul Lane 6. 10 ul |
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 (Cat. No.: 04543-51), Chemi-Lumi One L (Cat. No.: 07880-70) |
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Product | Storage | Cat.No. | PKG Size | Price | |
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Blocking One-P | 2-8°C | 05999-84 | 200 ml | 52.00 | Buy |