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Product: RayBio® Phosphorylation ELISA Kits
      Introduction
  Highlight
  How it works
  Contents of kit
  Price
  Representative Result
  List of Products
  Reference List 		  
Introduction
RayBio®  Human PhosphoELISA (Enzyme-Linked Immunosorbent Assay) kit is a very rapid, convenient and sensitive assay kit that can monitor the activation or function of important biological pathways in cell lysates. By determining phosphorylated protein in your experimental model system, you can verify pathway activation in your cell lysates. You can simultaneously measure numerous different cell lysates without spending excess time and effort in performing a Western Blot analysis.

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.
 
Highlight
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
How it works

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
Price
Product Species Description Raybiotech_Catalog Size Price
Phospho-Akt (Ser473) Human/ Mouse/ Rat Phospho-Akt (Ser473) for cell lysate PEL-Akt-S473-001 96 wells (12 strips x 8 wells) $428.00
Phospho-Akt (Ser473) and pan Akt Human/ Mouse/ Rat Phospho-Akt (Ser473) and pan Akt for cell lysate PEL-Akt-S473-002 96 wells (12 strips x 8 wells) $468.00
Phospho-EGFR (Tyr992) Human Phospho-EGFR (Tyr992) for cell lysate PEL-EGFR-Y992-001 96 wells (12 strips x 8 wells) $428.00
Phospho-EGFR (Tyr992) and pan EGFR Human Phospho-EGFR (Tyr992) and pan EGFR for cell lysate PEL-EGFR-Y992-002 96 wells (12 strips x 8 wells) $468.00
Phospho-EGFR (Tyr1068) Human Phospho-EGFR (Tyr1068) for cell lysate PEL-EGFR-Y1068-001 96 wells (12 strips x 8 wells) $428.00
Phospho-EGFR (Tyr1068) and pan EGFR Human Phospho-EGFR (Tyr1068) and pan EGFR for cell lysate PEL-EGFR-Y1068-002 96 wells (12 strips x 8 wells) $468.00
Phospho-EGFR (Tyr1045) Human Phospho-EGFR (Tyr1045) for cell lysate PEL-EGFR-Y1045-001 96 wells (12 strips x 8 wells) $428.00
Phospho-EGFR (Tyr1045) and pan EGFR Human Phospho-EGFR (Tyr1045) and pan EGFR for cell lysate PEL-EGFR-Y1045-002 96 wells (12 strips x 8 wells) $468.00
Phospho-EGFR (Ser1070) Human Phospho-EGFR (Ser1070) for cell lysate PEL-EGFR-S1070-001 96 wells (12 strips x 8 wells) $428.00
Phospho-EGFR (Ser1070) and pan EGFR Human Phospho-EGFR (Ser1070) and pan EGFR for cell lysate PEL-EGFR-S1070-002 96 wells (12 strips x 8 wells) $468.00
Phospho-EGFR (Tyr1086) Human Phospho-EGFR (Tyr1086) for cell lysate PEL-EGFR-Y1086-001 96 wells (12 strips x 8 wells) $428.00
Phospho-EGFR (Tyr1086) and pan EGFR Human Phospho-EGFR (Tyr1086) and pan EGFR for cell lysate PEL-EGFR-Y1086-002 96 wells (12 strips x 8 wells) $468.00
Phospho-Erk1Thr202 /Tyr204)/Erk2(Tyr185/187) Human/ Mouse/ Rat Phospho-Erk1Thr202 /Tyr204)/Erk2(Tyr185/187) for cell lysate PEL-Erk-T202-001 96 wells (12 strips x 8 wells) $428.00
Phospho-Erk1(Thr202 /Tyr204)/Erk2(Tyr185/187) and pan Erk1/2 Human/ Mouse/ Rat Phospho-Erk1(Thr202/Tyr204) /Erk2(Tyr185/187) and pan Erk1/2 for cell lysate PEL-Erk-T202-002 96 wells (12 strips x 8 wells) $468.00
Phospho-Mek1 (Ser217/221) Human/ Mouse/ Rat Phospho-Mek1 (Ser217/221) for cell lysate PEL-Mek-S217-001 96 wells (12 strips x 8 wells) $428.00
Phospho-Mek1 (Ser217/221) and pan Mek Human/ Mouse/ Rat Phospho-Mek1 (Ser217/221) and pan Mek for cell lysate PEL-Mek-S217-002 96 wells (12 strips x 8 wells) $468.00
Phospho-Met (Tyr1234) Human/ Mouse Phospho-Met (Tyr1234) for cell lysate PEL-Met-Y1234-001 96 wells (12 strips x 8 wells) $428.00
Phospho-Met (Tyr1234) and pan Mek Human/ Mouse Phospho-Met (Tyr1234) and pan Mek for cell lysate PEL-Met-Y1234-002 96 wells (12 strips x 8 wells) $468.00
Phospho-P38 alpha (Thr180/Tyr182) Human/ Mouse Phospho-P38 alpha (Thr180/Tyr182) for cell lysate PEL-P38-T180-001 96 wells (12 strips x 8 wells) $428.00
Phospho-P38 alpha (Thr180/Tyr182) and pan P38 Human/ Mouse Phospho-P38 alpha (Thr180/Tyr182) and pan P38 for cell lysate PEL-P38-T180-002 96 wells (12 strips x 8 wells) $468.00
Phospho-Stat1 (Tyr701) Human/ Mouse Phospho-Stat1 (Tyr701) for cell lysate PEL-Stat1-Y701-001 96 wells (12 strips x 8 wells) $428.00
Phospho-Stat1 (Tyr701) and pan Stat1 Human/ Mouse Phospho-Stat1 (Tyr701) and pan Stat1 for cell lysate PEL-Stat1-Y701-002 96 wells (12 strips x 8 wells) $468.00
Phospho-Stat1 (Ser727) Human/ Mouse Phospho-Stat1 (Ser727) for cell lysate PEL-Stat1-S727-001 96 wells (12 strips x 8 wells) $428.00
Phospho-Stat1 (Ser727) and pan Stat1 Human/ Mouse Phospho-Stat1 (Ser727) and pan Stat1 for cell lysate PEL-Stat1-S727-002 96 wells (12 strips x 8 wells) $468.00
Phospho-Stat3 (Tyr705) Human/ Mouse Phospho-Stat3 (Tyr705) for cell lysate PEL-Stat3-Y705-001 96 wells (12 strips x 8 wells) $428.00
Phospho-Stat3 (Tyr705) and pan Stat1 Human/ Mouse Phospho-Stat3 (Tyr705) and pan Stat1 for cell lysate PEL-Stat3-Y705-002 96 wells (12 strips x 8 wells) $468.00

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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.
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).

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Reference List
1. Profiling of cytokine expression by biotin-labeled-based proteinarrays. Ying Lin, Ruochun Huang, Lipai Chen et al. Proteomics.2003, 3: 1750-1757.
2. Proteomic profiling of the cancer microenvironment by antibody arrays. Vladimir Knezevic, Chidchanok Leethanakul, Verena E. Bichsel et al. Proteomics 2001, 1, 1271“1278
3. Antibody microarray profiling of human prostate cancer sera: Antibody screening and identification of potential biomarkersJeremy C. Miller, Heping Zhou, Joshua Kwekel, Robert Cavalloet al. The, Brian B. Haab. Proteomics. 2003, 3: 56-63.
4. Smad4 signalling in T cells is required for suppression of gastrointestinal cancer. Byung-Gyu Kim, Cuiling Li, WenhuiQiao, Mizuko Mamura, Barbara Kasperczak, Miriam Anver, Lawrence Wolfraim, Suntaek Hong, Elizabeth Mushinski, Michael Potter, Seong-Jin Kim, Xin-Yuan Fu, Chuxia Deng and John J. Letterio. Nature. 2006; Vol 441: 1015
5. Connexin suppresses human glioblastoma cell growth by down-regulation of monocyte chemotactic protein 1, as discoveredusing protein array technology. Ruochun Huang, Ying Lin, Cheng C. Wang, Jacob Gano, Biaoyang Lin, Qian Shi, AltonBoynton, Jocelyn Burke, and Ruo-Pan Huang. Cancer Res. 2002;62:2806-2812.
6. LPS induces the interaction of a transcription factor, LPS-induced TNF-a factor, and STAT6(B) with effects on multiple cytokines.Xiaoren Tang, Deborah Levy Marciano, Susan E. Leeman and Salomon Amar. PNAS. April 5, 2005 vol. 102 no. 14, 5132-5137
7. HIV-1-mediated apoptosis of neuronal cells: Proximal molecular mechanisms of HIV-1-induced encephalopathy. Yan Xu, JosephKulkoshy, Roger j. Pomerantz. PNAS. 2004 May 4, 2004 Vol. 101 No. 18.
8. A novel method for high-throughput protein profiling fromconditioned media and patient's sera. Ruo-Pan Huang, RuochunHuang, Yan Fan, and Ying Lin. Ana. Biochem. 2001;294(1):55-62.

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