Multicolor flow cytometric analysis of RT1B expression on rat splenocytes. Lewis rat splenocytes were stained with a PE Mouse Anti-Rat CD3 (Cat. No. 554833) and either Alexa Fluor® 647 Mouse IgG1, κ Isotype Control (Cat. No. 557714; left panel) or Alexa Fluor® 647 Mouse anti-Rat RT1B (Cat. No. 562223; right panel). Flow cytometric fluorescence dot plots showing RT1B expression (or Ig Isotype Control staining) versus CD3 were derived from gated events with the forward and side light-scattering characteristics of viable splenocytes. Flow cytometry was performed on a BD™ LSR II.
Multicolor flow cytometric analysis of RT1B expression on rat splenocytes. Lewis rat splenocytes were stained with a PE Mouse Anti-Rat CD3 (Cat. No. 554833) and either Alexa Fluor® 647 Mouse IgG1, κ Isotype Control (Cat. No. 557714; left panel) or Alexa Fluor® 647 Mouse anti-Rat RT1B (Cat. No. 562223; right panel). Flow cytometric fluorescence dot plots showing RT1B expression (or Ig Isotype Control staining) versus CD3 were derived from gated events with the forward and side light-scattering characteristics of viable splenocytes. Flow cytometry was performed on a BD™ LSR II.
Product Details
BD Pharmingen™
RT1-B; RT1 class II locus B
Rat (QC Testing)
Mouse BALB/c IgG1, κ
Ia-like Glycoproteins from Wistar Thymocytes
Flow cytometry (Routinely Tested)
0.2 mg/ml
AB_11153133
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
- 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.
- Alexa Fluor® is a registered trademark of Molecular Probes, Inc., Eugene, OR.
- Alexa Fluor® 647 fluorochrome emission is collected at the same instrument settings as for allophycocyanin (APC).
- 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.
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
Companion Products
Stain Buffer (FBS) RUO
Size
500 mL
Cat No.
554656
Stain Buffer (BSA) RUO
Size
500 mL
Cat No.
554657
Alexa Fluor® 647 Mouse IgG1 κ Isotype Control RUO
Size
100 Tests
Cat No.
557714
PE Mouse Anti-Rat CD3 RUO
Size
0.2 mg
Cat No.
554833
562223 Rev. 2
Antibody Details
OX-6
The OX-6 antibody specifically recognizes non-polymorphic determinants of the Rat MHC class II antigen, I-A equivalent. RT1B is found on peripheral B lymphocytes, thymic cortical epithelial and medullary reticular cells, small intestinal villus epithelium, epidermal Langerhans cells, dendritic cells, some tissue macrophage populations, peritoneal mast cells, and a subset of thymocytes, but not on peripheral T cells, erythrocytes, or microglia. The OX-6 mAb cross-reacts with mouse I-A[k] and I-A[s] alloantigens and with a major subset of splenocytes from NOD (I-A[g7]) mice.
562223 Rev. 2
Format Details
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 520-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
562223 Rev.2
Citations & References
Development References (10)
-
Chen-Woan M, Delaney CP, Fournier V, et al. In vitro characterization of rat bone marrow-derived dendritic cells and their precursors. J Leukoc Biol. 1996; 59(2):196-207. (Biology). View Reference
-
Damoiseaux JG, Yagita H, Okumura K, van Breda Vriesman PJ. Costimulatory molecules CD80 and CD86 in the rat; tissue distribution and expression by antigen-presenting cells. J Leukoc Biol. 1998; 64(6):803-809. (Biology). View Reference
-
Dick AD, Ford AL, Forrester JV, Sedgwick JD. Flow cytometric identification of a minority population of MHC class II positive cells in the normal rat retina distinct from CD45lowCD11b/c+CD4low parenchymal microglia. J Leukoc Biol. 1995; 79(9):834-840. (Biology). View Reference
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Fox CC, Jewell SD, Whitacre CC. Rat peritoneal mast cells present antigen to a PPD-specific T cell line. Cell Immunol. 1994; 158(1):253-264. (Biology). View Reference
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Fukumoto T, McMaster WR, Williams AF, et al. Mouse monoclonal antibodies against rat major histocompatibility antigens. Two Ia antigens and expression of Ia and class I antigens in rat thymus. Eur J Immunol. 1982; 12:237-243. (Biology). View Reference
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Mayrhofer G, Pugh CW, Barclay AN. The distribution, ontogeny and origin in the rat of Ia-positive cells with dendritic morphology and of Ia antigen in epithelia, with special reference to the intestine. Eur J Immunol. 1983; 13(2):112-122. (Biology). View Reference
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McMaster WR, Williams AF et al. Identification of Ia glycoproteins in rat thymus and purification from rat spleen.. Eur J Immunol. 1979; 9:426-433. (Immunogen). View Reference
-
Neiss U, Becker D, Knop J, Reske K. Modulation of MHC class II determinants on rat Langerhans cells during short term culture. Adv Exp Med Biol. 1993; 329:29-34. (Clone-specific). View Reference
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Nelson DJ, McMenamin C, McWilliam AS, Brenan M, Holt PG. Development of the airway intraepithelial dendritic cell network in the rat from class II major histocompatibility (Ia)-negative precursors: differential regulation of Ia expression at different levels of the respiratory tract. J Exp Med. 1994; 179(1):203-212. (Biology). View Reference
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Xia WJ, Schneeberger EE, McCarthy K, Kradin RL. Accessory cells of the lung. II. Ia+ pulmonary dendritic cells display cell surface antigen heterogeneity. Am J Respir Cell Mol Biol. 1991; 5(3):276-283. (Biology). View Reference
562223 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.
