Immunohistofluorescent analysis of CD3 expression by cells within B56BL/6 mouse spleen (left panel). A mouse spleen cryosection (5 µm) was fixed with BD Cytofix™ Fixation Buffer (Cat. No. 554655), blocked with 5% goat serum and 1% BSA diluted in 1x PBS, and stained with Alexa Fluor® 647 Rat Anti-Mouse CD3 Molecular Complex antibody (Cat. No. 557869, pseudo-colored green), BD Horizon™ BV421 Rat Anti-Mouse CD11b antibody (Cat. No. 562605, pseudo-colored red), and Alexa Fluor® 488 Rat Anti-Mouse CD45R/B220 antibody (Cat. No. 557669, pseudo-colored blue). Images were captured on a standard epifluorescence microscope. Original magnification, 20x. CD3 expression in spleen and thymus (middle and right panel). CD57BL/6 splenocytes were simultaneously stained with Alexa Fluor® 647 mAb 17A2 (Cat. No. 557869) and PE rat anti-mouse CD19 mAb 1D3 (Cat. No. 557399, middle panel). C57BL/6 thymocytes were stained with either Alexa Fluor® 647-conjugated mAb 17A2 (Cat. No. 557869, right panel, filled histogram) or Alexa Fluor® 647-conjugated rat IgG2b, κ isotype control mAb A95-1 (Cat. No. 557691) (right panel, empty histogram). Flow cytometry was performed on a BD FACSCalibur™ flow cytometry system.
Immunohistofluorescent analysis of CD3 expression by cells within B56BL/6 mouse spleen (left panel). A mouse spleen cryosection (5 µm) was fixed with BD Cytofix™ Fixation Buffer (Cat. No. 554655), blocked with 5% goat serum and 1% BSA diluted in 1x PBS, and stained with Alexa Fluor® 647 Rat Anti-Mouse CD3 Molecular Complex antibody (Cat. No. 557869, pseudo-colored green), BD Horizon™ BV421 Rat Anti-Mouse CD11b antibody (Cat. No. 562605, pseudo-colored red), and Alexa Fluor® 488 Rat Anti-Mouse CD45R/B220 antibody (Cat. No. 557669, pseudo-colored blue). Images were captured on a standard epifluorescence microscope. Original magnification, 20x. CD3 expression in spleen and thymus (middle and right panel). CD57BL/6 splenocytes were simultaneously stained with Alexa Fluor® 647 mAb 17A2 (Cat. No. 557869) and PE rat anti-mouse CD19 mAb 1D3 (Cat. No. 557399, middle panel). C57BL/6 thymocytes were stained with either Alexa Fluor® 647-conjugated mAb 17A2 (Cat. No. 557869, right panel, filled histogram) or Alexa Fluor® 647-conjugated rat IgG2b, κ isotype control mAb A95-1 (Cat. No. 557691) (right panel, empty histogram). Flow cytometry was performed on a BD FACSCalibur™ flow cytometry system.
Product Details
BD Pharmingen™
Mouse (QC Testing)
Rat SD, also known as Sprague-Dawley (outbred) IgG2b, κ
γδ TCR-positive T-T hybridoma D1
Flow cytometry (Routinely Tested), Immunofluorescence (Tested During Development)
0.2 mg/ml
12501
AB_396912
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.
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- 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.
- Alexa Fluor® is a registered trademark of Molecular Probes, Inc., Eugene, OR.
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
Companion Products
Alexa Fluor® 647 Rat IgG2b, κ Isotype Control RUO
Size
0.1 mg
Cat No.
557691
PE Rat Anti-Mouse CD19 RUO
Size
0.1 mg
Cat No.
557399
557869 Rev. 6
Antibody Details
17A2
The 17A2 monoclonal antibody specifically binds to the T-cell receptor-associated CD3 complex that is expressed on many thymocytes and mature T lymphocytes. Plate-bound 17A2 antibody has been reported to induce IL-2 production by cultured T cells in the absence of accessory cells. The binding of the 17A2 antibody to T cells can be blocked by the anti-CD3e mAb 145-2C11 (Cat. No. 557306/553058/550275). This suggests that the 17A2 antibody recognizes an epitope of the CD3 epsilon chain. In vivo treatment with 17A2 antibody has been reported to partially deplete T lymphocytes and temporarily down-modulates CD3 expression on T cells.
557869 Rev. 6
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 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
557869 Rev.6
Citations & References
View product citations for antibody "557869" on CiteAb
Development References (2)
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Miescher GC, Schreyer M, MacDonald HR. Production and characterization of a rat monoclonal antibody against the murine CD3 molecular complex. Immunol Lett. 1989; 23(2):113-118. (Immunogen: Cytotoxicity, Functional assay, Immunohistochemistry, Immunoprecipitation, Stimulation). View Reference
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Mysliwietz J, Thierfelder S. Antilymphocytic antibodies and marrow transplantation. XII. Suppression of graft-versus-host disease by T-cell-modulating and depleting antimouse CD3 antibody is most effective when preinjected in the marrow recipient. Blood. 1992; 80(10):2661-2667. (Clone-specific: Depletion). View Reference
557869 Rev. 6
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.