Phosphorylation ELISA Kits

Semi-Quantitative, Sandwich ELISAs for Detection of Phosphorylated Proteins in Cell Lysates

RayBio® Phosphorylation ELISA kits are rapid, convenient and sensitive assays that monitor the activation or function of important biological pathways in cell lysate samples. By determining phosphorylated protein in experimental model systems, researchers can verify pathway activation and simultaneously measure numerous different cell lysates without spending excess time and effort in performing Western Blot analyses.

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Phospho-ELISA kit


  • Simultaneously measure Phosphorylated protein and pan protein in one experiment (for normalization purpose)
  • Screen numerous different cell lysates without performing a Western Blot analysis
  • Minimal hands-on time, convenient, and non-radioactive material
  • High sensitivity
  • High quality
  • Cost-effective
  • Complete kit with pre-coated 96 well plate

Contents of Kit

  • 96 wells (12 strips x 8 wells) Coated With Antibody
  • Wash Buffer Concentrate (20x)
  • Positive Control
  • Assay Diluent
  • Detection Antibody
  • Secondary Antibody or HRP Streptavidin
  • TMB One-Step Substrate Reagent
  • Stop Solution
  • Cell Lysis Buffer

How It Works

Phospho-ELISA - How it works

This Sandwich ELISA kit is an in vitro enzyme-linked immunosorbent assay for the measurement of phosphorylated protein. An specific capture antibody (anti-pan-antibody or anti-phosphorylated antibody) has been coated onto a 96-well plate. Samples are pipetted into the wells and target protein present in a sample is bound to the wells by the immobilized antibody. The wells are washed and anti-phosphorylated antibody or anti-pan-antibody is used to detect target protein. After washing away unbound antibody, secondary antibody is pipetted to the wells. The wells are again washed, a TMB substrate solution is added to the wells and color develops in proportion to the amount of target bound. The Stop Solution changes the color from blue to yellow, and the intensity of the color is measured at 450 nm.

GSK3b Positive Control

Representative Results

1. Recombinant Human EGF Stimulation of A431 Cell Lines:

A431 cells were treated or untreated with 100 ng/ml recombinant human EGF for 10 min. Cell lysates were analyzed using this phosphoELISA and Western Blot.

Phospho ELISA

2. Sensitivity:

The A431 cells were treated with 100 ng/mL recombinant human EGF for 20 minutes to induce phosphorylation of EGF R. Serial dilutions of lysates were analyzed in this ELISA and by Western blot. Immunoblots were incubated with anti-phospho-EGFR (Tyr 1086).

Phospho ELISA Sensitivity

Why RayBiotech ELISA Kits?

  • We manufacture, not resell, ELISA kits. Right here in the USA.
  • Our ELISAs are ISO certified and validated for exceptional reproducibility. Read more about our quality control processes.
  • All RayBio® ELISAs are backed by our 90-day 100% satisfaction guarantee.
  • Discounts and incentives available for publications citing our products, and for product reviews.


  1. Al-Gayyar M., et al.  Protective effects of arjunolic acid against cardiac toxicity induced by oral sodium nitrite: Effects on cytokine balance and apoptosis. Life Sciences Available online 23 July 2014. [view publication]
    Product: PEL-JNK-T183-T
    Sample Type: Tissue Lysate
  2. Correale J., Farez M., Ysrraelit M. Role of prolactin in B cell regulation in multiple sclerosis. Journal of Neuroimmunology Volume 269, Issue 1 , Pages 76-86, 15 April 2014 [view publication]
    Products: PEL-Stat1-Y, PEL-Stat3-Y, PEL-Stat5-Y
    Sample Type: Cell Lysate
  3. Elamin E., et al. Ethanol Impairs Intestinal Barrier Function in Humans through Mitogen Activated Protein Kinase Signaling: A Combined In Vivo and In Vitro Approach. PLOS One, September 16, 2014 DOI: 10.1371/journal.pone.0107421 [view publication]
    Product: PEL-MAPK-SK
    Sample Type: Tissue Lysate
  4. Katsila T., et al. Circulating pEGFR is a Candidate Response Biomarker of Cetuximab Therapy in Colorectal Cancer. Clinical Cancer Research, October 2014, epub ahead of print. DOI: 10.1158/1078-0432.CCR-14-0361. [view publication]
    Product: PEL-EGFR-Y1068
    Sample Type: Plasma
  5. Tukaj S., Gruner D., Tukaj C., et al. Calcitriol exerts anti-inflammatory effects in keratinocytes treated with autoantibodies from a patient with bullous pemphigoid. J Eur Acad Dermatol Venereol. 2015 Jan 22. doi: 10.1111/jdv.12929 [view publication]
    Product: PEL-STAT3-Y705-T
    Sample Type: Cell Lysate
  6. Wang N., Rayes R., Elahi S., et al. The IGF-Trap: Novel Inhibitor of Carcinoma Growth and Metastasis. Mol Cancer Ther. 2015 Apr;14(4):982-93. doi: 10.1158/1535-7163.MCT-14-0751. [view publication]
    Product: PEL-AKT-S473-T
    Sample Type: Cell Lysate
  7. Di Grazia A., Cappiello F., Imanishi A., et al. The Frog Skin-Derived Antimicrobial Peptide Esculentin-1a(1-21)NH2 Promotes the Migration of Human HaCaT Keratinocytes in an EGF Receptor-Dependent Manner: A Novel Promoter of Human Skin Wound Healing? PLoS One. 2015 Jun 12;10(6):e0128663. doi: 10.1371/journal.pone.0128663 [view publication]
    Product: PEL-STAT3-Y705
    Sample Type: Cell Lysate
  8. Al-Maghrebi, May, et al. "Lutein modulates transcription dysregulation of adhesion molecules and spermatogenesis transcription factors induced by testicular ischemia reperfusion injury: it could be SAFE." Naunyn-Schmiedeberg's archives of pharmacology 389.5 (2016): 539-551. [view publication]
    Products: PEL-JAK1-Y1022-T, PEL-Stat3-Y705-T
    Sample Type: Tissue Lysate

More references are listed per product on the individual product pages.

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