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      Alexa Fluor™ 647 Rabbit Anti-Human HK1
      Alexa Fluor™ 647 Rabbit Anti-Human HK1
      Flow cytometric analysis of HK1 expression in HEK293T cells. Cells from the Human HEK293T (Embryonic Kidney, ATCC® CRL-1573™) cell line were fixed with BD Cytofix™ Fixation Buffer (Cat. No. 554655) and permeabilized with Perm/Wash Buffer (Cat. No. 554723). Cells were then stained with either Alexa Fluor™ 647 Rabbit IgG Isotype Control (Cat. No. 569345; dashed line histogram) or Alexa Fluor™ 647 Rabbit Anti-Human HK1 antibody (Cat. No. 569319; solid line histogram) at 0.125 µg/test. The fluorescence histogram showing the expression of HK1 (or Ig Isotype control staining) was derived from gated events with the forward and side light-scatter characteristics of intact cells. Flow cytometry and data analysis were performed using a BD LSRFortessa™ X-20 Cell Analyzer System and FlowJo™ software. Data shown on this Technical Data Sheet are not lot specific.
      Alexa Fluor™ 647 Rabbit Anti-Human HK1
      Flow cytometric analysis of HK1 expression in HEK293T cells. Cells from the Human HEK293T (Embryonic Kidney, ATCC® CRL-1573™) cell line were fixed with BD Cytofix™ Fixation Buffer (Cat. No. 554655) and permeabilized with Perm/Wash Buffer (Cat. No. 554723). Cells were then stained with either Alexa Fluor™ 647 Rabbit IgG Isotype Control (Cat. No. 569345; dashed line histogram) or Alexa Fluor™ 647 Rabbit Anti-Human HK1 antibody (Cat. No. 569319; solid line histogram) at 0.125 µg/test. The fluorescence histogram showing the expression of HK1 (or Ig Isotype control staining) was derived from gated events with the forward and side light-scatter characteristics of intact cells. Flow cytometry and data analysis were performed using a BD LSRFortessa™ X-20 Cell Analyzer System and FlowJo™ software. Data shown on this Technical Data Sheet are not lot specific.
      Product Details
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      BD Pharmingen™
      HK; hexokinase type I; hexokinase-1
      Human (QC Testing), Mouse (Reported)
      Rabbit IgG
      Human Hexokinase 1
      Intracellular staining (flow cytometry) (Routinely Tested)
      0.2 mg/ml
      Aqueous buffered solution containing ≤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 the dye under optimum conditions and unreacted dye was removed.
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      Recommended Assay Procedures

      BD® CompBeads can be used as surrogates to assess fluorescence spillover (compensation).  When fluorochrome conjugated antibodies are bound to BD® CompBeads, they have spectral properties very similar to cells.   However, for some fluorochromes there can be small differences in spectral emissions compared to cells, resulting in spillover values that differ when compared to biological controls.  It is strongly recommended that when using a reagent for the first time, users compare the spillover on cell and BD® CompBeads to ensure that BD® CompBeads are appropriate for your specific cellular application.

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      Product Notices

      1. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
      2. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
      3. An isotype control should be used at the same concentration as the antibody of interest.
      4. Alexa Fluor® 647 fluorochrome emission is collected at the same instrument settings as for allophycocyanin (APC).
      5. 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.
      6. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
      7. This product is provided under an intellectual property license between Life Technologies Corporation and BD Businesses. The purchase of this product conveys to the buyer the non-transferable right to use the purchased amount of the product and components of the product in research conducted by the buyer (whether the buyer is an academic or for-profit entity). The buyer cannot sell or otherwise transfer (a) this product (b) its components or (c) materials made using this product or its components to a third party or otherwise use this product or its components or materials made using this product or its components for Commercial Purposes. Commercial Purposes means any activity by a party for consideration and may include, but is not limited to: (1) use of the product or its components in manufacturing; (2) use of the product or its components to provide a service, information, or data; (3) use of the product or its components for therapeutic, diagnostic or prophylactic purposes; or (4) resale of the product or its components, whether or not such product or its components are resold for use in research. For information on purchasing a license to this product for any other use, contact Life Technologies Corporation, Cell Analysis Business Unit Business Development, 29851 Willow Creek Road, Eugene, OR 97402, USA, Tel: (541) 465-8300. Fax: (541) 335-0504.
      8. This rabbit monoclonal (RabMabTM) product is, in part, provided under an intellectual property license from Abcam. The purchase of this product conveys to the buyer the non-transferable right to use the purchased product and components of the product for buyer’s internal research use only (whether the buyer is an academic or for-profit entity) and buyer shall not be permitted to reverse engineer or otherwise perform any compositional, structural, functional or other analyses directed to learning the methodology, formulae, sequences, process, make-up or production of this product (or any portion thereof). The sale of this product is expressly conditioned on the buyer not using the product or its components, or any materials made using the product or its components, in any activity to generate revenue, which may include, but is not limited to use of the product or its components: (i) in manufacturing; (ii) to provide a service, information, or data in return for payment (iii) for therapeutic, diagnostic or prophylactic purposes; or (iv) for resale, regardless of whether they are sold for use in research. If you, as the buyer, do not agree to these conditions, please leave the product unopened and return the contents of this package to BD Biosciences, 10975 Torreyana Road, San Diego, CA 92121. For information on purchasing a license to this product for purposes other than research, contact Abcam at partnerships@abcam.com.
      9. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
      10. Alexa Fluor™ is a trademark of Life Technologies Corporation.
      11. For U.S. patents that may apply, see bd.com/patents.
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      569319 Rev. 2
      Antibody Details
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      EPR10134(B)

      The EPR10134(B) recombinant monoclonal antibody specifically recognizes Hexokinase 1 (HK1), which is also known as HK, Hexokinase-1 or Hexokinase type I. HKI is a ~ 100 kDa glycolytic enzyme that is encoded by the HKI gene, which belongs to the hexokinase family. This isoenzyme contains a N-terminal regulatory domain and a C-terminal catalytic domain fused into a single polypeptide chain in the outer surface of mitochondria. HK1 binds to mitochondria through the interaction with porin, the protein associated with the modulation of cellular permeability. In glucose metabolism, HKI is one of four mammalian transferase enzymes that catalyze the ATP-dependent phosphorylation of hexoses in cells and tissues. HKI is highly expressed in erythrocytes, platelets, brain, kidney, and heart. This isozyme functions primarily in a catabolic role by mediating the initial step of glycolysis to form glucose 6-phosphate (G6P). Cytosolic HKI increases macrophage-mediated inflammatory cytokine production and acts as a pattern recognition receptor for bacterial peptidoglycan in innate immunity.

      569319 Rev. 2
      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 670-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
      569319 Rev.2
      Citations & References
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      View product citations for antibody "569319" on CiteAb

      Development References (9)

      1. Ahl PJ, Hopkins RA, Xiang WW, et al. Met-Flow, a strategy for single-cell metabolic analysis highlights dynamic changes in immune subpopulations.. Communications Biology. 2020; 3(1):1-15. (Clone-specific: Flow cytometry). View Reference
      2. Aleshin AE, Zeng C, Bartunik HD, Fromm HJ, Honzatko RB. Regulation of hexokinase I: crystal structure of recombinant human brain hexokinase complexed with glucose and phosphate.. J Mol Biol. 1998; 282(2):345-57. (Biology). View Reference
      3. Aleshin AE, Zeng C, Bourenkov GP, Bartunik HD, Fromm HJ, Honzatko RB. The mechanism of regulation of hexokinase: new insights from the crystal structure of recombinant human brain hexokinase complexed with glucose and glucose-6-phosphate.. Structure. 1998; 6(1):39-50. (Biology). View Reference
      4. De Jesus A, Keyhani-Nejad F, Pusec CM, et al. Hexokinase 1 cellular localization regulates the metabolic fate of glucose.. Mol Cell. 2022; 82(7):1261-1277.e9. (Biology). View Reference
      5. Gamradt S, Hasselmann H, Taenzer A, et al. Reduced mitochondrial respiration in T cells of patients with major depressive disorder.. iScience. 2021; 24(11):103312. (Clone-specific: Flow cytometry). View Reference
      6. Magnani M, Bianchi M, Casabianca A, et al. A recombinant human 'mini'-hexokinase is catalytically active and regulated by hexose 6-phosphates.. Biochem J. 1992; 285 ( Pt 1):193-9. (Biology). View Reference
      7. Reinfeld BI, Madden MZ, Wolf MM, et al. Cell-programmed nutrient partitioning in the tumour microenvironment.. Nature. 2021; 593(7858):282-288. (Clone-specific: Flow cytometry). View Reference
      8. Wilson JE. Isozymes of mammalian hexokinase: structure, subcellular localization and metabolic function.. J Exp Biol. 2003; 206(Pt 12):2049-57. (Biology). View Reference
      9. Wolf AJ, Reyes CN, Liang W, et al. Hexokinase Is an Innate Immune Receptor for the Detection of Bacterial Peptidoglycan.. Cell. 2016; 166(3):624-636. (Biology). View Reference
      View All (9) View Less
      569319 Rev. 2

       

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      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.