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 제품안내  면역학   Cell Signaling  Phosphate Assay

Phosphoprotein Detection & Enrichment

항체를 사용하지 않고 phosphoprotein을 detection 할수 있는 kit (pIMAGO)와 Phosphopeptide를 농축(enrichment) 하는 kit (PolyMAC)을 소개합니다.

Tymora-Analytical
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특징

 pIMAGO Phosphoprotein Detection
 
  • pIMAGO는 water-soluble, globular nanopolymers로 phosphate group에 잘 붙습니다.
  • Phosphoreylated Amino acid를 detection하는 항체가 필요없습니다.
  • 종(species), organism에 상관없이 모든 phosphoprotein을 잡을 수 있습니다.
  • 항체를 사용하지 않아서 항상 일관된 결과를 얻을수 있습니다.
  • 값비싸고 핸들링이 어려운 방사성동위원소를 사용하지 않아서 안전합니다.
  • 특이도가 높습니다. (즉 phosphoprotein에만 반응합니다.)
  • 민감도가 높습니다. (즉 적은 양의 phosphoprotein도 잘 검출됩니다.)
  • 인단백질과 결합력이 좋아서, washing 혹은 reprobing 중에 떨어져 나갈 염려가 없습니다.
  • Western blot 및 ELISA 실험에 항체 대신 사용할 수 있습니다.
  • 항체와 함께 이용시 total 및 phosphorylated protein을 함께 측정할 수 있는 multiplexing 효과를 얻을수 있습니다.
  • 적용 : in vitro kinase/phosphatase assay, Kinase activity 및 inhibitor screening
  • Kit 구성품 :
    - pIMAGO-biotin reagent
    - Avidin-HRP 혹은 Avidin-Fluor
    - 각종 buffer (blocking buffer, pIMAGO buffer, washing buffer)
    - control phosphoprotein
    - 96 well plate (ECL용 Kit의 경우)

    ◆ 적용방법

 PolyMAC Phosphopeptide Enrichment
  • PolyMAC (Polymer-based Metal-ion Affinity Capture)는 MASS 분석을 위한 peptide 샘플에서 phosphopeptide를 분리하는데 사용되며, In-Solution에서 진행되므로 reproducibility, selectivity, sensitivity를 높혀줍니다.
  • Phosphopeptide recovery가 좋습니다.
  • Phosphopeptide와 PolyMAC의 결합단계와 Bead를 이용한 분리단계로 진행됩니다.
  • Bead의 경우, magnetic bead 혹은 agarose bead를 이용할 수 있습니다.
  • Kit 구성품 :
    - PolyMAC-Ti reagent
    - capture beads
    - loading buffer, washing buffer, capture buffer and elution buffer
    - Agarose based kit의 경우, re-usable spin columns 제공

   ◆ 비디오 프로토콜 보기

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   ◆ 사용한 논문 보기

  • Walls C., Iliuk A., Bai Y., Wang M., Tao WA, Zhang ZY. (2013). Phosphatase of regenerating liver 3 (PRL3) provokes a tyrosine phosphoproteome to drive prometastatic signal transduction. Mol. Cell. Proteomics. 12(12): 3759-77, http://www.ncbi.nlm.nih.gov/pubmed/24030100
  • Radu M., Rawat S., Beeser A., Iliuk A., Tao WA., Chernoff J. (2013). ArhGAP15, a Rac-specific GTPase activating protein, plays a dual role in inhibiting small GTPase signaling. Journal of Biological Chemistry. 288(29): 21117-25, http://www.ncbi.nlm.nih.gov/pubmed/23760270
  • Yu S., Huang H., Iliuk A., Wang W-H., Tao WA., Post C., Geahlen R. (2013). Syk Inhibits the Activity of Protein Kinase A by Phosphorylating Tyrosine 330 of the Catalytic Subunit. Journal of Biological Chemistry. 288(15): 10870-10881, http://www.ncbi.nlm.nih.gov/pubmed/23447535
  • Pan L, Iliuk A, Yu S, Geahlen RL, Tao WA. (2012). Multiplexed quantitation of protein expression and phosphorylation based on functionalized soluble nanopolymers. J Am Chem Soc. 134(44): 18201-4, http://www.ncbi.nlm.nih.gov/pubmed/23088311   Original pIMAGO-Fluor detection on microplate publication.
  • Iliuk A, Liu XS, Xue L, Liu X, Tao WA. (2012). Chemical visualization of phosphoproteomes on membrane. Mol. Cell. Proteomics. 11(9): 629-39, http://www.ncbi.nlm.nih.gov/pubmed/22593177   Original pIMAGO-biotin detection on Western Blot publication.
  • Xue L, Wang WH, Iliuk A, Hu L, Galan JA, Yu S, Hans M, Geahlen RL, Tao WA (2012). Sensitive kinase assay linked with phosphoproteomics for identifying direct kinase substrates. PNAS. 109(15): 5615-20, http://www.ncbi.nlm.nih.gov/pubmed/22451900
  • Iliuk A, Martinez J, Hall MC, Tao WA (2011). Phosphorylation assay based on functionalized soluble nanopolymer. Anal. Chem. 83(7): 2767-74, http://www.ncbi.nlm.nih.gov/pubmed/21395237  Original pIMAGO-biotin detection on microplate publication.
  • Iliuk AB, Martin VA, Alicie BM, Geahlen RL and Tao WA (2010). In-depth analyses of kinase-dependent tyrosine phosphoproteomes based on metal ion functionalized soluble nanopolymers. Mol. Cell. Proteomics. 9: 2162-2172, http://www.ncbi.nlm.nih.gov/pubmed/20562096  Original PolyMAC-Ti publication.
주문정보(Ordering Information)
CATNUM PRODUCT SIZE
BLOCK-WB-20010x Optimized Blocking Buffer for Western Blot - 200mL200 ml
BLOCK-WB-5010x Optimized Blocking Buffer for Western Blot - 50mL50 ml
BLOCK-PL-2501x Optimized Blocking Buffer for Microplate - 250mL250 ml
BLOCK-PL-501x Optimized Blocking Buffer for Microplate - 50mL50 ml
AGARB-100Agarose Capture Beads for PolyMAC Enrichment - 100 enrichments (5mL; used with spin columns)5 ml
AGARB-50Agarose Capture Beads for PolyMAC Enrichment - 50 enrichments (2.5mL; used with spin columns)2.5 ml
FL550-200Avidin-Fluor 550 - 200uL200 ul
FL550-2000Avidin-Fluor 550 - 2mL2 ml
FL550-500Avidin-Fluor 550 - 500uL500 ul
FL680-200Avidin-Fluor 680 - 200uL200 ul
FL680-2000Avidin-Fluor 680 - 2mL2 ml
FL680-500Avidin-Fluor 680 - 500uL500 ul
FL800-200Avidin-Fluor 800 - 200uL200 ul
FL800-2000Avidin-Fluor 800 - 2mL2 ml
FL800-500Avidin-Fluor 800 - 500uL500 ul
HRP-200Avidin-Peroxidase (avidin-HRP) - 200uL200 ul
HRP-2000Avidin-Peroxidase (avidin-HRP) - 2mL2 ml
HRP-500Avidin-Peroxidase (avidin-HRP) - 500uL500 ul
CNTRL-ELISAControl Phosphoprotein for ELISA (microplate detection)200 ul
CNTRL-WBControl Phosphoprotein for Western Blot (on-membrane detection)100 ul
PMGO-200Extra pIMAGO (reagent only) - 200uL200 ul
PMGO-500Extra pIMAGO (reagent only) - 500uL500 ul
702-1000Extra PolyMAC (reagent only) - 1mL (for 100 enrichments)1 ml
702-500Extra PolyMAC (reagent only) - 500uL (for 50 enrichments)500 ul
SPIN-10Extra Spin Columns for PolyMAC Agarose Capture - 10 spin columns (used with agarose beads)ea
SPIN-100Extra Spin Columns for PolyMAC Agarose Capture - 100 spin columns (used with agarose beads)ea
PLATE-20High Binding Capacity Clear 96-well Plates - 20 plates20 plates
PLATE-5High Binding Capacity Clear 96-well Plates - 5 plates5 plates
MAGB-100Magnetic Capture Beads for PolyMAC Enrichment - 100 enrichments (5mL)5 ml
MAGB-50Magnetic Capture Beads for PolyMAC Enrichment - 50 enrichments (2.5mL)2.5 ml
BIND-200Optimized On-plate Protein Binding Buffer - 200mL200 ml
BIND-50Optimized On-plate Protein Binding Buffer - 50mL50 ml
901-100pIMAGO Fluor 550 Phosphoprotein Detection on Microplate (complete kit) - 100 wells1 kit
901-400pIMAGO Fluor 550 Phosphoprotein Detection on Microplate (complete kit) - 400 wells1 kit
801-10pIMAGO Fluor 550 Phosphoprotein Detection on Western Blot (complete kit) - 10 mini-blots1 kit
801-40pIMAGO Fluor 550 Phosphoprotein Detection on Western Blot (complete kit) - 40 mini-blots1 kit
902-100pIMAGO Fluor 680 Phosphoprotein Detection on Microplate (complete kit) - 100 wells1 kit
902-400pIMAGO Fluor 680 Phosphoprotein Detection on Microplate (complete kit) - 400 wells1 kit
802-10pIMAGO Fluor 680 Phosphoprotein Detection on Western Blot (complete kit) - 10 mini-blots1 kit
802-40pIMAGO Fluor 680 Phosphoprotein Detection on Western Blot (complete kit) - 40 mini-blots1 kit
903-100pIMAGO Fluor 800 Phosphoprotein Detection on Microplate (complete kit) - 100 wells1 kit
903-400pIMAGO Fluor 800 Phosphoprotein Detection on Microplate (complete kit) - 400 wells1 kit
803-10pIMAGO Fluor 800 Phosphoprotein Detection on Western Blot (complete kit) - 10 mini-blots1 kit
803-40pIMAGO Fluor 800 Phosphoprotein Detection on Western Blot (complete kit) - 40 mini-blots1 kit
900-400pIMAGO HRP Colorimetric Phosphoprotein Detection on Microplate (complete kit) - 100 wells1 kit
900-100pIMAGO HRP Colorimetric Phosphoprotein Detection on Microplate (complete kit) - 100 wells1 kit
800-10pIMAGO HRP Phosphoprotein Detection on Western Blot (complete kit) - 10 mini-blots1 kit
800-40pIMAGO HRP Phosphoprotein Detection on Western Blot (complete kit) - 40 mini-blots1 kit
701-100PolyMAC-Ti Agarose Phosphopeptide Enrichment (complete kit) - 100 enrichments1 kit
701-50PolyMAC-Ti Agarose Phosphopeptide Enrichment (complete kit) - 50 enrichments1 kit
700-100PolyMAC-Ti Magnetic Phosphopeptide Enrichment (complete kit) - 100 enrichments1 kit
700-50PolyMAC-Ti Magnetic Phosphopeptide Enrichment (complete kit) - 50 enrichments1 kit