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      Alexa Fluor® 488 Mouse anti-CrkL (pY207)
      Alexa Fluor® 488 Mouse anti-CrkL (pY207)
      Analysis of CrkL (pY207) in human chronic myelogenous leukemia cell line. K562 cells (ATCC CCL-243, left panel) were either treated with the tyrosine kinase inhibitor, imatinib, at 25 μM (LC Laboratories, shaded histogram) for 2 hours at 37°C or untreated (open 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 at least 30 minutes, and then stained with Alexa Fluor® 488 Mouse anti-CrkL (pY207, Cat. No. 560789). Flow cytometry was performed on a BD FACSCanto™ II flow cytometry system. The specificity of mAb K30-391.50.80 was confirmed by western blot (right panel) using unconjugated antibody on lysates from control (lane 1) and imatinib-treated (lane 2) K562 cells. CrkL (pY207) is identified as a band of 39 kDa that has decreased intensity in the treated cells.
      Alexa Fluor® 488 Mouse anti-CrkL (pY207) Alexa Fluor® 488 Mouse anti-CrkL (pY207)
      Analysis of CrkL (pY207) in human chronic myelogenous leukemia cell line. K562 cells (ATCC CCL-243, left panel) were either treated with the tyrosine kinase inhibitor, imatinib, at 25 μM (LC Laboratories, shaded histogram) for 2 hours at 37°C or untreated (open 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 at least 30 minutes, and then stained with Alexa Fluor® 488 Mouse anti-CrkL (pY207, Cat. No. 560789). Flow cytometry was performed on a BD FACSCanto™ II flow cytometry system. The specificity of mAb K30-391.50.80 was confirmed by western blot (right panel) using unconjugated antibody on lysates from control (lane 1) and imatinib-treated (lane 2) K562 cells. CrkL (pY207) is identified as a band of 39 kDa that has decreased intensity in the treated cells.
      製品詳細
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      BD Phosflow™
      Human (QC Testing)
      Mouse BALB/c IgG2a, κ
      Phosphorylated Human, Mouse, and Rat CrkL Peptide
      Intracellular staining (flow cytometry) (Routinely Tested)
      20 µl
      AB_10529344
      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® 488 under optimum conditions, and unreacted Alexa Fluor® 488 was removed.
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      推奨アッセイ手順

      This Alexa Fluor® 488-conjugated antibody is suitable for intracellular staining of cell lines using BD Cytofix™ Fixation Buffer and BD Phosflow™ Perm Buffer III or IV. We have confirmed that the Alexa Fluor® 647 conjugate, Cat. No. 560790, is suitable for staining leukocytes from human peripheral blood and bone marrow.

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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. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
      3. 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.
      4. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
      5. Alexa Fluor® 488 fluorochrome emission is collected at the same instrument settings as for fluorescein isothiocyanate (FITC).
      6. 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.
      7. Alexa Fluor® is a registered trademark of Molecular Probes, Inc., Eugene, OR.
      8. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
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      560789 Rev. 2
      抗体の詳細
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      K30-391.50.80

      Crk-Like (CrkL) is an adaptor protein that is preferentially expressed in hematopoietic cells and is encoded by a gene that is homologous to the viral oncogene v-crk (chicken tumor virus no. 10 regulator of kinase). Its SH2 and SH3 domains bind to a variety of effector proteins, such as paxillin, p130Cas, c-Cbl, c-Abl, and C3G. These interactions are involved in the regulation of cellular migration, adhesion, and transformation. Tyrosine 207 (Y207) of CrkL is phosphorylated in hematopoietic cells that express the BCR-ABL fusion protein. This site may be a negative regulator of protein complex formation and biological activity.

      The K30-391.50.80 monoclonal antibody recognizes the phosphorylated Y207 of human CrkL.

      560789 Rev. 2
      フォーマットの詳細
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      Alexa Fluor™ 488
      Alexa Fluor™ 488 Dye is part of the BD blue family of dyes. This is a small organic fluorochrome with an excitation maximum (Ex Max) at 494-nm and an emission maximum (Em Max) at 517-nm. Alexa Fluor™ 488 is designed to be excited by the Blue laser (488 nm) and detected using an optical filter centered near 520-nm (e.g., a 530/30-nm bandpass filter). Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
      Alexa Fluor™ 488
      Blue 488 nm
      494 nm
      517 nm
      560789 Rev.2
      引用＆参考文献
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      Development References (3)

      1. Arai A, Nosaka Y, Kohsaka H, Miyasaka N, Miura O. CrkL activates integrin-mediated hematopoietic cell adhesion through the guanine nucleotide exchange factor C3G. Blood. 1999; 93(11):3713-3722. (Biology). View Reference
      2. Feller SM. Crk family adaptors-signalling complex formation and biological roles. Oncogene. 2001; 20:6348-6371. (Biology). View Reference
      3. Senechal K, Heaney C, Druker B, Sawyers CL. Structural requirements for function of the Crkl adapter protein in fibroblasts and hematopoietic cells. Mol Cell Biol. 1998; 18(9):5082-5090. (Biology). View Reference
      560789 Rev. 2

      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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