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Alexa Fluor® 647 Rat Anti-Mouse I-A/I-E

BD Pharmingen™ Alexa Fluor® 647 Rat Anti-Mouse I-A/I-E

Clone M5/114.15.2 (also known as M5/114)

(RUO)
Alexa Fluor® 647 Rat Anti-Mouse  I-A/I-E
Multicolor flow cytometric analysis of I-A/I-E MHC class II alloantigen expression on splenocytes from positive and negative mouse strains. Mouse spleen cells from either M5/114-negative SJL (Left Panel) or M5/114-positive BALB/c (Right Panel) mice were stained with Alexa Fluor® 647 Rat Anti-Mouse  I-A/I-E (Cat. No. 562367), PE Anti-Mouse CD45R/B220 (Cat. No. 553089/553090/561878) and PE Anti-Mouse CD11b (Cat. No. 553311/557397/561689) antibodies. Two-color flow cytometric dot plots showing the expression of I-A/I-E MHC class II alloantigens versus CD45R/B220 and CD11b were derived from gated events with the forward and side light-scatter characteristics of viable splenocytes. The M5/114 monoclonal antibody detects I-Ad and I-Ed MHC class II alloantigens that are expressed on both B cells and macrophages. Flow cytometry was performed using a BD™ LSR II Flow Cytometer System.
Multicolor flow cytometric analysis of I-A/I-E MHC class II alloantigen expression on splenocytes from positive and negative mouse strains. Mouse spleen cells from either M5/114-negative SJL (Left Panel) or M5/114-positive BALB/c (Right Panel) mice were stained with Alexa Fluor® 647 Rat Anti-Mouse  I-A/I-E (Cat. No. 562367), PE Anti-Mouse CD45R/B220 (Cat. No. 553089/553090/561878) and PE Anti-Mouse CD11b (Cat. No. 553311/557397/561689) antibodies. Two-color flow cytometric dot plots showing the expression of I-A/I-E MHC class II alloantigens versus CD45R/B220 and CD11b were derived from gated events with the forward and side light-scatter characteristics of viable splenocytes. The M5/114 monoclonal antibody detects I-Ad and I-Ed MHC class II alloantigens that are expressed on both B cells and macrophages. Flow cytometry was performed using a BD™ LSR II Flow Cytometer System.
Product Details
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BD Pharmingen™
H-2I; I-Ab, I-Ad, I-Aq, I-Ed, and I-Ek MHC class II alloAgs; Ia Ag; M5/114; MHC II
Mouse (QC Testing)
Rat BN x LEW IgG2b, κ
Activated C57BL/6 Mouse Spleen Cells
Flow cytometry (Routinely Tested)
0.2 mg/ml
111861
AB_11152078
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 Alexa Fluor® 647 under optimum conditions, and unreacted Alexa Fluor® 647 was removed.

Product Notices

  1. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
  2. An isotype control should be used at the same concentration as the antibody of interest.
  3. 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.
  4. Alexa Fluor® is a registered trademark of Molecular Probes, Inc., Eugene, OR.
  5. Alexa Fluor® 647 fluorochrome emission is collected at the same instrument settings as for allophycocyanin (APC).
  6. 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.
  7. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  8. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
562367 Rev. 2
Antibody Details
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M5/114.15.2

The M5/114.15.2 monoclonal antibody recognizes a polymorphic determinant shared by the I-A[b], I-A[d], I-A[q], I-E[d], and I-E[k] (but not I-A[f], I-A[k], or I-A[s]) MHC class II alloantigens that can be expressed by B cells, dendritic cells, monocytes, macrophages and activated T cells. It also reacts with cells from mice of the H-2[p] and H-2[r] haplotypes, and it is non-reactive with cells from NOD (H-2[g7]) mice. Flow cytometric analysis indicates that the M5/114.15.2 and 2G9 monoclonal antibodies have comparable reactivity on cells from mice with I-A[b], I-A[d], I-A[g7], I-A[q], I-E[d], and I-E[k] alloantigens.

562367 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
562367 Rev.2
Citations & References
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Development References (7)

  1. Bhattacharya A, Dorf ME, Springer TA. A shared alloantigenic determinant on Ia antigens encoded by the I-A and I-E subregions: evidence for I region gene duplication. J Immunol. 1981; 127(6):2488-2495. (Immunogen: Immunoprecipitation). View Reference
  2. Guo MW, Watanabe T, Mori E, Mori T. Molecular structure and function of CD4 on murine egg plasma membrane. Zygote. 1995; 3(1):65-73. (Clone-specific: Blocking). View Reference
  3. Hattori M, Buse JB, Jackson RA, et al. The NOD mouse: recessive diabetogenic gene in the major histocompatibility complex. Science. 1986; 231(4739):733-735. (Clone-specific: Flow cytometry). View Reference
  4. Nelson AJ, Hosier S, Brady W, Linsley PS, Farr AG. Medullary thymic epithelium expresses a ligand for CTLA4 in situ and in vitro. J Immunol. 1998; 151(5):2453-2461. (Clone-specific: Immunofluorescence, Immunohistochemistry). View Reference
  5. Shi Y, Kaliyaperumal A, Lu L, et al. Promiscuous presentation and recognition of nucleosomal autoepitopes in lupus: role of autoimmune T cell receptor alpha chain. J Exp Med. 1998; 187(3):367-378. (Clone-specific: Blocking). View Reference
  6. Viville S, Neefjes J, Lotteau V, et al. Mice lacking the MHC class II-associated invariant chain. Cell. 1993; 72(4):635-648. (Clone-specific: Flow cytometry, Immunofluorescence). View Reference
  7. Yamashita I, Nagata T, Tada T, Nakayama T. CD69 cell surface expression identifies developing thymocytes which audition for T cell antigen receptor-mediated positive selection. Int Immunol. 1993; 5(9):1139-1150. (Clone-specific: Blocking). View Reference
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562367 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.