Multiparameter flow cytometric analysis of CD35 expression on human peripheral blood leucocytes. Human whole blood was stained with either Alexa Fluor® 647 Mouse IgG1, κ Isotype Control (Cat. No. 557714; Left Panel) or Alexa Fluor® 647 Mouse Anti-Human CD35 antibody (Cat. No. 565329; Right Panel). The erythrocytes were lysed with BD FACS™ Lysing Solution (Cat. No.349202). Two-parameter flow cytometric contour plots showing the correlated expression of CD35 (or Ig Isotype control staining) versus side light-scatter (SSC-A) signals were derived from gated events with the forward and side light-scatter characteristics of intact leucocyte populations. Flow cytometric analysis was performed using a BD™ LSR II Flow Cytometer System.
Multiparameter flow cytometric analysis of CD35 expression on human peripheral blood leucocytes. Human whole blood was stained with either Alexa Fluor® 647 Mouse IgG1, κ Isotype Control (Cat. No. 557714; Left Panel) or Alexa Fluor® 647 Mouse Anti-Human CD35 antibody (Cat. No. 565329; Right Panel). The erythrocytes were lysed with BD FACS™ Lysing Solution (Cat. No.349202). Two-parameter flow cytometric contour plots showing the correlated expression of CD35 (or Ig Isotype control staining) versus side light-scatter (SSC-A) signals were derived from gated events with the forward and side light-scatter characteristics of intact leucocyte populations. Flow cytometric analysis was performed using a BD™ LSR II Flow Cytometer System.
Product Details
BD Pharmingen™
CR1; Complement receptor type 1; C3b/C4b receptor; C3BR; C4BR; KN
Human (QC Testing)
Mouse IgG1, κ
Human Cells of the Monocyte Lineage
Flow cytometry (Routinely Tested)
5 µl
III 204
1378
AB_2739186
Aqueous buffered solution containing BSA and ≤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
- This reagent has been pre-diluted for use at the recommended Volume per Test. We typically use 1 × 10^6 cells in a 100-µl experimental sample (a test).
- 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.
- Source of all serum proteins is from USDA inspected abattoirs located in the United States.
- 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.
- 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
Lysing Solution 10X Concentrate IVD
Size
100 mL
Cat No.
349202
Lysing Buffer RUO
Size
100 mL
Cat No.
555899
Alexa Fluor® 647 Mouse IgG1 κ Isotype Control RUO
Size
100 Tests
Cat No.
557714
565329 Rev. 1
Antibody Details
E11
The E11 monoclonal antibody specifically binds to CD35. CD35 is also known as Complement receptor type 1 (CR1), C3b/C4b receptor, C3BR, C4BR, Immune adherence receptor, or KN. CD35 is a type I transmembrane glycoprotein that exists in four allelic forms of 160, 190, 220 and 250 kDa. CD35 serves as a receptor for complement fragments C3b, iC3b, C3dg, C4b, iC3, and iC4. It enhances phagocytosis by neutrophils and monocytes and regulates complement activation. It is expressed on erythrocytes, granulocytes, monocytes, B cells, and some dendritic cells, T cells, and NK cells. It binds complement components C3b and C4b, mediating. This antibody cannot inhibit the phagocytic capacity of granulocytes. The CD35 antibody is useful in studies of cells that express complement receptors.
565329 Rev. 1
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
565329 Rev.1
Citations & References
Development References (6)
-
Barclay NA, Brown MH, Birkeland ML, et al, ed. The Leukocyte Antigen FactsBook. San Diego, CA: Academic Press; 1997.
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Dougherty GJ, Selvendran Y, Murdoch S, Palmer DG, Hogg N. The human mononuclear phagocyte high-affinity Fc receptor, FcRI, defined by a monoclonal antibody, 10.1. Eur J Immunol. 1987; 17(10):1453-1459. (Biology). View Reference
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Hogg N, Ross GD, Jones DB, Slusarenko M, Walport MJ, Lachmann PJ. Identification of an anti-monocyte monoclonal antibody that is specific for membrane complement receptor type one (CR1). Eur J Immunol. 1984; 14(3):236-243. (Immunogen: Blocking, Fluorescence microscopy, Functional assay, Immunofluorescence, Immunohistochemistry, Immunoprecipitation, Inhibition, Radioimmunoassay). View Reference
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Lin G-X, Yang X, Hollemweguer E, et al. Cross-reactivity of CD antibodies in eight animal species. In: Mason D. David Mason .. et al., ed. Leucocyte typing VII : white cell differentiation antigens : proceedings of the Seventh International Workshop and Conference held in Harrogate, United Kingdom. Oxford: Oxford University Press; 2002:519-523.
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Nickells M, Hauhart R, Krych M, et al. Mapping epitopes for 20 monoclonal antibodies to CR1. Clin Exp Immunol. 1998; 112(1):27-33. (Clone-specific: Dot Blot, ELISA). View Reference
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Schlossman SF. Stuart F. Schlossman .. et al., ed. Leucocyte typing V : white cell differentiation antigens : proceedings of the fifth international workshop and conference held in Boston, USA, 3-7 November, 1993. Oxford: Oxford University Press; 1995.
565329 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.
