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Blocking Reagent for Immunoassays with Phospho-Specific Antibodies: Blocking One-P

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


Western Blotting with phospho-specific antibody

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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  5. Kato, Masaya, et al. "Dual‐specificity phosphatase 6 plays a critical role in the maintenance of a cancer stem‐like cell phenotype in human endometrial cancer." International Journal of Cancer (2020).

  6. Imai, Takahiko, et al. "Levetiracetam, an antiepileptic drug has neuroprotective effects on intracranial hemorrhage injury." Neuroscience 431 (2020): 25-33.

  7. Honda, Mizuki, et al. "High resolution spatial transcriptome analysis by photo-isolation chemistry." bioRxiv (2020).

  8. Khafagy, El-Sayed, et al. "Systemic and brain delivery of leptin via intranasal coadministration with cell-penetrating peptides and its therapeutic potential for obesity." Journal of Controlled Release (2020).

  9. Kusumoto, Junya, et al. "OPN4 belongs to the photosensitive system of the human skin." Genes to Cells 25.3 (2020): 215-225.

Ordering Information

Product Cat.No. Storage PKG Size Price(US$)  
Blocking One-P 05999-84 2-8°C 200 ml 52.00 Buy