Multiparameter flow cytometric analysis of Integrin αE2 (CD103) expression on GM-CSF stimulated splenic leucocytes. Lewis rat Splenocytes were cultured overnight with Recombinant Rat GM-CSF (Cat. No. 555111). The cells were harvested and then pre-incubated with Purified Mouse Anti-Rat CD32 (Cat. No. 550270/550271). The cells were then stained with either Alexa Fluor® 647 Mouse IgG1 κ Isotype Control (Cat. No. 557714, left panel) or Alexa Fluor® 647 Mouse Anti-Rat Integrin αE2 (CD103) (Cat. No. 565286; right panel). Two-parameter flow cytometric contour plots showing the correlated expression of Integrin αE2 (CD103) [or Ig Isotype control staining] versus side light-scatter (SSC-A) signals were derived from gated events with the forward and side-light scattering characteristics of viable leucocytes. Flow cytometric analysis was performed on a BD™ LSR II.
Multiparameter flow cytometric analysis of Integrin αE2 (CD103) expression on GM-CSF stimulated splenic leucocytes. Lewis rat Splenocytes were cultured overnight with Recombinant Rat GM-CSF (Cat. No. 555111). The cells were harvested and then pre-incubated with Purified Mouse Anti-Rat CD32 (Cat. No. 550270/550271). The cells were then stained with either Alexa Fluor® 647 Mouse IgG1 κ Isotype Control (Cat. No. 557714, left panel) or Alexa Fluor® 647 Mouse Anti-Rat Integrin αE2 (CD103) (Cat. No. 565286; right panel). Two-parameter flow cytometric contour plots showing the correlated expression of Integrin αE2 (CD103) [or Ig Isotype control staining] versus side light-scatter (SSC-A) signals were derived from gated events with the forward and side-light scattering characteristics of viable leucocytes. Flow cytometric analysis was performed on a BD™ LSR II.
Product Details
BD Pharmingen™
Integrin alpha E; Itgae; Integrin αE; CD103
Rat (QC Testing)
Mouse BALB/c IgG1, κ
Density gradient-enriched PVG rat thoracic-duct dendritic cells
Flow cytometry (Routinely Tested)
0.2 mg/ml
AB_2739156
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.
- 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.
- 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).
- 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
Purified Mouse Anti-Rat CD32 RUO
Size
0.1 mg
Cat No.
550270
Purified Mouse Anti-Rat CD32 RUO
Size
0.5 mg
Cat No.
550271
Recombinant Rat GM-CSF RUO
Size
5 µg
Cat No.
555111
565286 Rev. 2
Antibody Details
OX-62
The OX-62 monoclonal antibody recognizes an antigen expressed on dendritic cells, dendritic epidermal T cells (but not RT1B+ Langerhans cells), and intestinal intraepithelial lymphocytes of normal rats and on CD3+ lymphocytes in the spleen and cervical lymph nodes of athymic nude rats. The antigen can be detected by flow cytometric or immunocytochemical analysis of leucocytes and by immunohistochemical staining of epidermal sheets, whole mounts of neural tissue, and frozen sections of lymphoid and nonlymphoid organs. The OX-62 monoclonal antibody immunoprecipitates rat Integrin αE2, a molecule that functions in leucocyte adhesion and homing. Sequence analysis reveals that this molecule is homologous to mouse and human CD103 (Integrin αIEL or αE chain).
565286 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
565286 Rev.2
Citations & References
View product citations for antibody "565286" on CiteAb
Development References (7)
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Brenan M, Puklavec M. The MRC OX-62 antigen: a useful marker in the purification of rat veiled cells with the biochemical properties of an integrin. J Exp Med. 1992; 175(6):1457-1465. (Immunogen: Immunofluorescence, Immunohistochemistry, Immunoprecipitation, Western blot). View Reference
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Brenan M, Rees DJ. Sequence analysis of rat integrin alpha E1 and alpha E2 subunits: tissue expression reveals phenotypic similarities between intraepithelial lymphocytes and dendritic cells in lymph. Eur J Immunol. 1997; 27(11):3070-3079. (Clone-specific: Immunofluorescence, Immunohistochemistry, Western blot). View Reference
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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. (Clone-specific: Immunocytochemistry (cytospins)). View Reference
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Gieseler R, Hoffmann PR, Kuhn R, et al. Enrichment and characterization of dendritic cells from rat renal mesangium. Scand J Immunol. 1997; 46(6):587-596. (Clone-specific: Immunohistochemistry). View Reference
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McMenamin PG. Distribution and phenotype of dendritic cells and resident tissue macrophages in the dura mater, leptomeninges, and choroid plexus of the rat brain as demonstrated in wholemount preparations. J Comp Neurol. 1999; 405(4):553-562. (Clone-specific: Immunofluorescence, Immunohistochemistry). 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. (Clone-specific: Immunohistochemistry). View Reference
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Penfield JG, Wang Y, Li S, et al. Transplant surgery injury recruits recipient MHC class II-positive leukocytes into the kidney. Kidney Int. 1999; 56(5):1759-1769. (Clone-specific: Immunohistochemistry). View Reference
565286 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.