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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