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      Alexa Fluor® 647 Mouse anti-ERK2
      Alexa Fluor® 647 Mouse anti-ERK2
      Analysis of ERK2 in human epithelioid carcinoma.  HeLa S3 cells (ATCC CCL 2.2) were either transfected with ERK2 RNAi (open histogram) or untransfected (shaded histogram).  The cells were fixed (BD Cytofix™ buffer, Cat. No. 554655) for 10 minutes at 37°C, then permeabilized (BD Phosflow™ Perm Buffer III, Cat. No. 558050) on ice for 30 minutes, and then stained with Alexa Fluor® 647 Mouse anti-ERK2 (Cat. No. 558526).  Down-regulation of ERK2 expression is evident in the RNAi-transfected cells.  Flow cytometry was performed on a BD FACSArray™ bioanalyzer system.
      Alexa Fluor® 647 Mouse anti-ERK2
      Analysis of ERK2 in human epithelioid carcinoma.  HeLa S3 cells (ATCC CCL 2.2) were either transfected with ERK2 RNAi (open histogram) or untransfected (shaded histogram).  The cells were fixed (BD Cytofix™ buffer, Cat. No. 554655) for 10 minutes at 37°C, then permeabilized (BD Phosflow™ Perm Buffer III, Cat. No. 558050) on ice for 30 minutes, and then stained with Alexa Fluor® 647 Mouse anti-ERK2 (Cat. No. 558526).  Down-regulation of ERK2 expression is evident in the RNAi-transfected cells.  Flow cytometry was performed on a BD FACSArray™ bioanalyzer system.
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      BD Phosflow™
      Human, Mouse (QC Testing)
      Mouse IgG1, κ
      Human ERK2 Synthetic Peptide
      Intracellular staining (flow cytometry) (Routinely Tested)
      20 µl
      AB_647125
      Aqueous buffered solution containing BSA and ≤0.09% sodium azide.
      RUO


      Preparation And Storage

      Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze. The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography. The antibody was conjugated to Alexa Fluor® 647 under optimum conditions, and unreacted Alexa Fluor® 647 was removed.
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      Product Notices

      1. This reagent has been pre-diluted for use at the recommended Volume per Test. We typically use 1 × 10^6 cells in a 100-µl experimental sample (a test).
      2. An isotype control should be used at the same concentration as the antibody of interest.
      3. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
      4. Caution: Sodium azide yields highly toxic hydrazoic acid under acidic conditions. Dilute azide compounds in running water before discarding to avoid accumulation of potentially explosive deposits in plumbing.
      5. Alexa Fluor® 647 fluorochrome emission is collected at the same instrument settings as for allophycocyanin (APC).
      6. Alexa Fluor® is a registered trademark of Molecular Probes, Inc., Eugene, OR.
      7. The Alexa Fluor®, Pacific Blue™, and Cascade Blue® dye antibody conjugates in this product are sold under license from Molecular Probes, Inc. for research use only, excluding use in combination with microarrays, or as analyte specific reagents. The Alexa Fluor® dyes (except for Alexa Fluor® 430), Pacific Blue™ dye, and Cascade Blue® dye are covered by pending and issued patents.
      8. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
      9. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
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      558526 Rev. 3
      Antibody Details
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      G263-7

      The members of the Mitogen-Activated Protein Kinase (MAPK) family are components of a key signal transduction cascade that links events at the cell surface to responses in the nucleus. The signaling cascade is found in species as varied as yeast and humans, with many of the proteins being well conserved.  In mammals the most widely studied members of the cascade are the Extracellular signal-Regulated Kinases, ERK1 (p44 MAPK) and ERK2 (p42 MAPK). ERK1 and ERK2 share 85% homology and are activated by extracellular signals such as growth factors, hormones, and phorbol esters. Activation occurs through a series of phosphorylations by kinases activating other kinases and eventually leading to phosphorylation of the ERKs. Growth factor stimulation leads to activation of Ras and Raf, leading to phosphorylation of MEK1 (MAPK/ERK kinase) which, in turn, activates the ERKs via dual phosphorylation. Once activated, the ERKs phosphorylate other cytoplasmic signaling molecules (protein kinases and phosphatases), cell-surface receptors, microtubule-associated proteins, and transcription factors in the nucleus. Thus, the active ERK has myriad downstream effectors that implicate it in the control of cell proliferation and differentiation, as well as regulation of the cytoskeleton.  Furthermore, studies have shown that elevated ERK activity is associated with some cancers. The G263-7 recognizes ERK2.  It does not cross-react with ERK1. Clone G263-7 was originally characterized in human (A431) and mouse (NIH/3T3) cells. A human ERK2 synthetic peptide was used as immunogen.

      558526 Rev. 3
      Format Details
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      Alexa Fluor™ 647
      Alexa Fluor™ 647 Dye is part of the BD red family of dyes. This is a small organic fluorochrome with an excitation maximum (Ex Max) at 653-nm and an emission maximum (Em Max) at 669-nm. Alexa Fluor 647 is designed to be excited by the Red laser (627-640 nm) and detected using an optical filter centered near 520-nm (e.g., a 660/20 nm bandpass filter). Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
      Alexa Fluor™ 647
      Red 627-640 nm
      653 nm
      669 nm
      558526 Rev.3
      Citations & References
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      Development References (9)

      1. Cobb MH, Boulton TG, Robbins DJ. Extracellular signal-regulated kinases: ERKs in progress. Cell Regul. 1991; 2:965-978. (Biology). View Reference
      2. Davis RJ. The mitogen-activated protein kinase signal transduction pathway. J Biol Chem. 1993; 268(20):14553-14556. (Biology). View Reference
      3. Fang JY, Richardson BC. The MAPK signalling pathways and colorectal cancer. Lancet Oncol. 2005; 6(5):322-327. (Biology). View Reference
      4. Kim SC, Hahn JS, Min YH, Yoo NC, Ko YW, Lee WJ. Constitutive activation of extracellular signal-regulated kinase in human acute leukemias: combined role of activation of MEK, hyperexpression of extracellular signal-regulated kinase, and downregulation of a phosphatase, PAC1. Blood. 1999; 93(11):3893-3899. (Biology). View Reference
      5. Reiser V, Ammerer G, Ruis H. Nucleocytoplasmic traffic of MAP kinases. Gene Expr. 1999; 7(4-6):247-254. (Biology). View Reference
      6. Sivaraman VS, Wang H, Nuovo GJ, Malbon CC. Hyperexpression of mitogen-activated protein kinase in human breast cancer. J Clin Invest. 1997; 99(7):1478-1483. (Biology). View Reference
      7. Stupack DG, Cho SY, Klemke RL. Molecular signaling mechanisms of cell migration and invasion. Immunol Res. 2000; 21(2-3):83-88. (Biology). View Reference
      8. Sweatt JD. The neuronal MAP kinase cascade: a biochemical signal integration system subserving synaptic plasticity and memory. J Neurochem. 2001; 76:1-10. (Biology).
      9. Treisman R. Regulation of transcription by MAP kinase cascades. Curr Opin Cell Biol. 1996; 8:205-215. (Biology). View Reference
      View All (9) View Less
      558526 Rev. 3

      Please refer to Support Documents for Quality Certificates


      Global - Refer to manufacturer's instructions for use and related User Manuals and Technical data sheets before using this products as described


      Comparisons, where applicable, are made against older BD Technology, manual methods or are general performance claims.  Comparisons are not made against non-BD technologies, unless otherwise noted.

      For Research Use Only. Not for use in diagnostic or therapeutic procedures.

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