BD Pharmingen™ PE-Cy7 Hamster Anti-Mouse CD11c (Integrin αX)
Clone N418 (RUO)
Multicolor flow cytometric analysis of CD11c (Integrin αX) expression on viable Mouse splenic leukocytes. In order to reduce nonspecific background staining, BD Pharmingen™ MonoBlock™ Leukocyte Staining Buffer (Cat. No. 570002) was added and mixed with C57BL/6 Mouse splenocytes followed by preincubation with Purified Rat Anti-Mouse CD16/CD32 antibody (Mouse BD Fc Block™) [Cat. No. 553142]. The splenic leukocytes were then stained with BD Horizon™ BV650 Rat Anti-Mouse I-A/I-E antibody (Cat. No. 563415) and with either PE-Cy7 Hamster IgG1, κ Isotype Control (Cat. No. 552811; Left Plot; used as a representative Hamster IgG Isotype Control) or PE-Cy7 Hamster Anti-Mouse CD11c (Integrin αX) antibody (Cat. No. 570804/570880; Right Plot) at 1 µg/test. DAPI Solution (Cat. No. 564907) was added to cells right before analysis. The bivariate pseudocolor density plot showing the correlated expression of CD11c (Integrin αX) [or Ig Isotype control staining] versus I-A/I-E was derived from gated events with the forward and side light-scatter characteristics of viable (DAPI-negative) splenic leukocytes. Flow cytometry and data analysis were performed using a BD LSRFortessa™ X-20 Cell Analyzer System and FlowJo™ Software. Data shown on this Technical Data Sheet are not lot specific.
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Multicolor flow cytometric analysis of CD11c (Integrin αX) expression on viable Mouse splenic leukocytes. In order to reduce nonspecific background staining, BD Pharmingen™ MonoBlock™ Leukocyte Staining Buffer (Cat. No. 570002) was added and mixed with C57BL/6 Mouse splenocytes followed by preincubation with Purified Rat Anti-Mouse CD16/CD32 antibody (Mouse BD Fc Block™) [Cat. No. 553142]. The splenic leukocytes were then stained with BD Horizon™ BV650 Rat Anti-Mouse I-A/I-E antibody (Cat. No. 563415) and with either PE-Cy7 Hamster IgG1, κ Isotype Control (Cat. No. 552811; Left Plot; used as a representative Hamster IgG Isotype Control) or PE-Cy7 Hamster Anti-Mouse CD11c (Integrin αX) antibody (Cat. No. 570804/570880; Right Plot) at 1 µg/test. DAPI Solution (Cat. No. 564907) was added to cells right before analysis. The bivariate pseudocolor density plot showing the correlated expression of CD11c (Integrin αX) [or Ig Isotype control staining] versus I-A/I-E was derived from gated events with the forward and side light-scatter characteristics of viable (DAPI-negative) splenic leukocytes. Flow cytometry and data analysis were performed using a BD LSRFortessa™ X-20 Cell Analyzer System and FlowJo™ Software. Data shown on this Technical Data Sheet are not lot specific.
Product Details
BD Pharmingen™
Itgax; Integrin alpha-X; Integrin αX; ITAX; CR4; Complement Receptor 4
Mouse (QC Testing)
Armenian Hamster IgG2
Mouse Dendritic Cells
Flow cytometry (Routinely Tested)
0.2 mg/ml
Aqueous buffered solution containing ≤0.09% sodium azide.
RUO
Preparation And Storage
The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography. The antibody was conjugated to the dye under optimum conditions and unconjugated antibody and free dye were removed. Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze.
Recommended Assay Procedures
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
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
- 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.
- Since applications vary, each investigator should titrate the reagent to obtain optimal results.
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- Warning: Some APC-Cy7 and PE-Cy7 conjugates show changes in their emission spectrum with prolonged exposure to formaldehyde. If you are unable to analyze fixed samples within four hours, we recommend that you use BD™ Stabilizing Fixative (Cat. No. 338036).
- An isotype control should be used at the same concentration as the antibody of interest.
- PE-Cy7 is a tandem fluorochrome composed of R-phycoerythrin (PE), which is excited by 488-nm light and serves as an energy donor, coupled to the cyanine dye Cy7, which acts as an energy acceptor and fluoresces maximally at 780 nm. PE-Cy7 tandem fluorochrome emission is collected in a detector for fluorescence wavelengths of 750 nm and higher. Although every effort is made to minimize the lot-to-lot variation in the efficiency of the fluorochrome energy transfer, differences in the residual emission from PE may be observed. Therefore, we recommend that individual compensation controls be performed for every PE-Cy7 conjugate. PE-Cy7 is optimized for use with a single argon ion laser emitting 488-nm light, and there is no significant overlap between PE-Cy7 and FITC emission spectra. When using dual-laser cytometers, which may directly excite both PE and Cy7, we recommend the use of cross-beam compensation during data acquisition or software compensation during data analysis.
- Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
- Please observe the following precautions: We recommend that special precautions be taken (such as wrapping vials, tubes, or racks in aluminum foil) to protect exposure of conjugated reagents, including cells stained with those reagents, to any room illumination. Absorption of visible light can significantly affect the emission spectra and quantum yield of tandem fluorochrome conjugates.
- Cy is a trademark of Global Life Sciences Solutions Germany GmbH or an affiliate doing business as Cytiva.
- For U.S. patents that may apply, see bd.com/patents.
Companion Products
Stain Buffer (FBS) RUO
Size
500 mL
Cat No.
554656
Stain Buffer (BSA) RUO
Size
500 mL
Cat No.
554657
MonoBlock™ Leukocyte Staining Buffer RUO
Size
500 Tests
Cat No.
570002
Purified Rat Anti-Mouse CD16/CD32 (Mouse BD Fc Block™) RUO
Size
0.5 mg
Cat No.
553142
PE-Cy™7 Hamster IgG1, κ Isotype Control RUO
Size
0.1 mg
Cat No.
552811
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BV650 Rat Anti-Mouse I-A/I-E RUO
Size
50 µg
Cat No.
563415
570804 Rev. 1
Antibody Details
N418
The N418 monoclonal antibody specifically binds to CD11c, the Integrin alpha X (Integrin αX, Itgax) chain of the heterodimeric gp150, 95 (CD11c/CD18, αXβ2) integrin that forms the complement receptor 4 (CR4). CD11c is a150 kDa type I transmembrane glycoprotein that is expressed on dendritic cells, CD4-CD8+ intestinal intraepithelial lymphocytes (IEL), and some NK cells and T cells. CD11c expression is upregulated on IEL and T cells following activation. Cells of the monocyte/macrophage lineage have been reported to express low levels of CD11c. The CD11c/CD18 integrin can bind to several ligands including iC3b, fibrinogen, and CD54. This integrin reportedly plays important roles in phagocytosis and in mediating cellular interactions during inflammation.
570804 Rev. 1
Format Details
PE-Cy7
PE-Cy7 dye is a part of the BD PE family of dyes. This tandem fluorochrome is comprised of a R-Phycoerythrin (PE) donor that has excitation maxima (Ex Max) of 496-nm and 566-nm and an acceptor dye, Cy™7, with an emission maximum (Em Max) at 781-nm. PE can be excited by the Blue (488-nm), Green (532-nm) and yellow-green (561-nm) lasers and detected using an optical filter centered near 781 nm (e.g., a 760/60-nm bandpass filter). The donor dye can be excited by the Blue (488-nm), Green (532-nm) and yellow-green (561-nm) lasers and the acceptor dye can be excited by the Red (627–640-nm) laser resulting in cross-laser excitation and fluorescence spillover. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
PE-Cy7
Yellow-Green 561 nm
496 nm, 566 nm
781 nm
570804 Rev.1
Citations & References
View product citations for antibody "570804" on CiteAb
Development References (4)
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Crowley MT, Inaba K, Witmer-Pack MD, Gezelter S, Steinman RM. Use of the fluorescence activated cell sorter to enrich dendritic cells from mouse spleen. J Immunol Methods. 1990; 133(1):55-66. (Clone-specific: Flow cytometry, Fluorescence activated cell sorting). View Reference
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Metlay JP, Witmer-Pack MD, Agger R, Crowley MT, Lawless D, Steinman RM. The distinct leukocyte integrins of mouse spleen dendritic cells as identified with new hamster monoclonal antibodies. J Exp Med. 1990; 171(5):1753-1771. (Immunogen: Flow cytometry, Immunohistochemistry, Immunoprecipitation). View Reference
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Orozco SL, Daniels BP, Yatim N, et al. RIPK3 Activation Leads to Cytokine Synthesis that Continues after Loss of Cell Membrane Integrity.. Cell Rep. 2019; 28(9):2275-2287.e5. (Clone-specific: Flow cytometry). View Reference
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Sadhu C, Ting HJ, Lipsky B, et al. CD11c/CD18: novel ligands and a role in delayed-type hypersensitivity. J Leukoc Biol. 2007; 81(6):1395-1403. (Clone-specific: Blocking). View Reference
570804 Rev. 1
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.