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Alexa Fluor™ 647 Rat Anti-Human HLA-B
Alexa Fluor™ 647 Rat Anti-Human HLA-B

Multiparameter flow cytometric analysis of HLA-B expression on Human peripheral blood leukocyte populations.  Human whole blood was first treated with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899) to lyse erythrocytes, washed, and then preincubated with BD Pharmingen™ Human BD Fc Block™ (Cat. No. 564219/564220). The leukocytes were then stained with either Alexa Fluor™ 647 Rat IgG1, κ Isotype Control (Cat. No. 557731; Left Plot) or Alexa Fluor™ 647 Rat Anti-Human HLA-B antibody (Cat. No. 569662/569663; Right Plot). The bivariate pseudocolor density plot showing the correlated expression of HLA-B (or Ig Isotype control staining) versus side light-scatter (SSC-A) signals was derived from gated events with the forward and side-light scatter characteristics of viable leukocytes. Flow cytometry and data analysis were performed using a BD LSRFortessa™ X-20 Cell Analyzer System and FlowJo™ software.

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569662
¥66,000
EA (1 Each)
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製品詳細
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BD Pharmingen™
HLA-B; HLAB
Human (QC Testing)
Rat IgG1, κ
Normal Human Lymphocytes
Flow cytometry (Routinely Tested)
5 µl/test
3106
AB_3685212
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 antibody was conjugated to the dye under optimum conditions and unreacted dye was removed. The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography.

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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.

Product Notices

  1. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
  2. 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).
  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. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
  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.

データシート

569662 Rev. 2
抗体の詳細
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YTH 76.3.rMAb

The YTH 76.3rMAb is a recombinant monoclonal antibody that specifically recognizes a monomorphic epitope on the extracellular region of the Human Leukocyte Antigen-B (HLA-B) heavy chain. This major histocompatibility complex (MHC) class I antigen is comprised of a polymorphic HLA-B heavy chain, a ~45 kDa type I transmembrane glycoprotein encoded by HLA-B, that is noncovalently associated with the invariant ~12 kDa Beta-2 (β2)-microglobulin light chain encoded by B2M. Hundreds of HLA-B alleles have been reported. HLA-B is expressed on most nucleated cells and leucocytes including specialized antigen presenting cells such as thymic epithelial cells and dendritic cells. HLA-B plays a major role in the MHC-restricted presentation of small peptides, including those derived from self or foreign antigens, that can be bound by TCR expressed on CD8+ thymocytes or T cells leading to either immune tolerance, the maturation of naïve CD8+ T cells, or the activation and differentiation of CD8+ cytotoxic effector T cells or memory CD8+ T cells. HLA-B also functions as a ligand for regulatory CD8 coreceptor molecules and some CD158 molecules that serve as regulatory MHC class I antigen receptors expressed by CD8+ T cells and NK cells. The YTH 76.3rMAb is derived from the hybridoma clone YTH 76.3, with Rat IgG2a, κ isotype. YTH 76.3rMAb has variable region sequences from the original YTH 76.3 clone appended to Rat IgG1 and kappa constant region sequences.

569662 Rev. 2
フォーマットの詳細
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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
569662 Rev.2
引用&参考文献
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Development References (6)

  1. Burrone OR, Kefford RF, Gilmore D, Milstein C. Stimulation of HLA-A,B,C by IFN-alpha. The derivation of Molt 4 variants and the differential expression of HLA-A,B,C subsets.. EMBO J. 1985; 4(11):2855-60. (Clone-specific: Flow cytometry). View Reference
  2. Hale G, Clark M, Waldmann H. Therapeutic potential of rat monoclonal antibodies: isotype specificity of antibody-dependent cell-mediated cytotoxicity with human lymphocytes.. J Immunol. 1985; 134(5):3056-61. (Immunogen). View Reference
  3. Margulies DH, Natarajan K, Rossjohn J, McCluskey J. Major Histocompatibility Complex and Its Proteins. In: Paul WE. Paul WE, ed. Fundamental Immunology 7th Edition. Philadelphia: Lippincott Williams & Wilkins; 2013:487-523.
  4. Rojas RE, Balaji KN, Subramanian A, Boom WH. Regulation of human CD4(+) alphabeta T-cell-receptor-positive (TCR(+)) and gammadelta TCR(+) T-cell responses to Mycobacterium tuberculosis by interleukin-10 and transforming growth factor beta.. Infect Immun. 1999; 67(12):6461-72. (Clone-specific: Flow cytometry). View Reference
  5. Schmidt H, Gekeler V, Haas H, et al. Differential regulation of HLA class I genes by interferon.. Immunogenetics. 1990; 31(4):245-52. (Biology). View Reference
  6. Serrano A, Tanzarella S, Lionello I, et al. Rexpression of HLA class I antigens and restoration of antigen-specific CTL response in melanoma cells following 5-aza-2'-deoxycytidine treatment.. Int J Cancer. 2001; 94(2):243-51. (Clone-specific: Flow cytometry). View Reference
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569662 Rev. 2

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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.