Preparation And Storage
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 cells and BD® CompBeads to ensure that BD® CompBeads are appropriate for your specific cellular application.
For optimal and reproducible results, BD Horizon Brilliant Stain Buffer should be used anytime BD Horizon Brilliant dyes are used in a multicolor flow cytometry panel. Fluorescent dye interactions may cause staining artifacts which may affect data interpretation. The BD Horizon Brilliant Stain Buffer was designed to minimize these interactions. When BD Horizon Brilliant Stain Buffer is used in the multicolor panel, it should also be used in the corresponding compensation controls for all dyes to achieve the most accurate compensation. For the most accurate compensation, compensation controls created with either cells or beads should be exposed to BD Horizon Brilliant Stain Buffer for the same length of time as the corresponding multicolor panel. More information can be found in the Technical Data Sheet of the BD Horizon Brilliant Stain Buffer (Cat. No. 563794/566349) or the BD Horizon Brilliant Stain Buffer Plus (Cat. No. 566385).
Note: When using high concentrations of antibody, background binding of this dye to erythroid cell subsets (mature erythrocytes and precursors) has been observed. For researchers studying these cell populations, or in cases where light scatter gating does not adequately exclude these cells from the analysis, this background may be an important factor to consider when selecting reagents for panel(s).
Product Notices
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
- Since applications vary, each investigator should titrate the reagent to obtain optimal results.
- An isotype control should be used at the same concentration as the antibody of interest.
- 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.
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- Human donor specific background has been observed in relation to the presence of anti-polyethylene glycol (PEG) antibodies, developed as a result of certain vaccines containing PEG, including some COVID-19 vaccines. We recommend use of BD Horizon Brilliant™ Stain Buffer in your experiments to help mitigate potential background. For more information visit https://www.bdbiosciences.com/en-us/support/product-notices.
- Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
- 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.
- For U.S. patents that may apply, see bd.com/patents.
Companion Products
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.
Development References (9)
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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
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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
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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
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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
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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
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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
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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
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Wilson JE. Isozymes of mammalian hexokinase: structure, subcellular localization and metabolic function.. J Exp Biol. 2003; 206(Pt 12):2049-57. (Biology). View Reference
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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
Please refer to Support Documents for Quality Certificates
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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.