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R718 Rat Anti-Mouse CD13
R718 Rat Anti-Mouse CD13

Two-color flow cytometric analysis of CD13 expression on mouse splenocytes. Mouse splenic leucocytes were preincubated with Purified Rat Anti-Mouse CD16/CD32 antibody (Mouse BD Fc Block™) (Cat. No. 553141/553142). The cells were then stained with PE Rat Anti-Mouse CD11b antibody (Cat. No. 553311/557397/561689) and either BD Horizon™ R718 Rat IgG1, κ Isotype Control (Cat. No. 566948; Left Plot) or BD Horizon™ R718 Rat Anti-Mouse CD13 antibody (Cat. No. 566982; Right Plot) at 2 µg/test. A bivariate pseudocolor density plot showing the correlated expression of CD13 (or Ig Isotype control staining) versus CD11b was derived from gated events with the forward and side light-scatter characteristics of viable leucocytes. Flow cytometry and data analysis were performed using a BD LSRFortessa™ Flow Cytometer System and FlowJo™ software. Data shown on this Technical Data Sheet are not lot specific.

Two-color flow cytometric analysis of CD13 expression on mouse splenocytes. Mouse splenic leucocytes were preincubated with Purified Rat Anti-Mouse CD16/CD32 antibody (Mouse BD Fc Block™) (Cat. No. 553141/553142). The cells were then stained with PE Rat Anti-Mouse CD11b antibody (Cat. No. 553311/557397/561689) and either BD Horizon™ R718 Rat IgG1, κ Isotype Control (Cat. No. 566948; Left Plot) or BD Horizon™ R718 Rat Anti-Mouse CD13 antibody (Cat. No. 566982; Right Plot) at 2 µg/test. A bivariate pseudocolor density plot showing the correlated expression of CD13 (or Ig Isotype control staining) versus CD11b was derived from gated events with the forward and side light-scatter characteristics of viable leucocytes. Flow cytometry and data analysis were performed using a BD LSRFortessa™ Flow Cytometer System and FlowJo™ software. Data shown on this Technical Data Sheet are not lot specific.

Product Details
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BD Horizon™
Anpep; Alanyl aminopeptidase; Aminopeptidase N; AP-N; mAPN; Lap-1
Mouse (QC Testing)
Rat LEW, also known as Lewis IgG1
Crude Aminopeptidase N Preparation from BALB/c Mouse Intestinal Epithelium
Flow cytometry (Routinely Tested)
0.2 mg/ml
AB_2869987
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 the dye under optimum conditions and unreacted dye was removed.

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 cells and BD CompBead to ensure that BD CompBeads are appropriate for your specific cellular application.

Product Notices

  1. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
  2. An isotype control should be used at the same concentration as the antibody of interest.
  3. 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.
  4. Alexa Fluor® is a registered trademark of Life Technologies Corporation.
  5. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
  6. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
  7. This product is provided under an Agreement between BIOTIUM and BD Biosciences. This product, and only in the amount purchased by buyer, may be used solely for buyer’s own internal research, in a manner consistent with the accompanying product literature. No other right to use, sell or otherwise transfer (a) this product, or (b) its components is hereby granted expressly, by implication or by estoppel. This product is for research use only. Diagnostic uses require a separate license from Biotium, Inc. For information on purchasing a license to this product including for purposes other than research, contact Biotium, Inc., 3159 Corporate Place, Hayward, CA 94545, Tel: (510) 265-1027. Fax: (510) 265-1352. Email: btinfo@biotium.com.
566982 Rev. 3
Antibody Details
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R3-242

The R3-242 monoclonal antibody specifically binds to aminopeptidase N (APN), which has been reported to be the mouse CD13 homologue. APN is expressed in a variety of mammalian tissues, including intestinal and renal epithelial brush border, endothelium, liver, lung, and brain. In lymphoid tissues, R3-242 mAb is reported to detect APN on monocytes, macrophages, mast cells, dendritic cells, and B lymphocytes, but not on T cells, and thymocytes. However, the reported reactivity with B cells may be non-specific, via binding to the Fcγ receptors (CD16/CD32). This observation is in agreement with reports using different antibodies to mouse APN. APN may play a role in antigen presentation in the immune system. R3-242 mAb does not inhibit the enzymatic activity of APN.

The antibody was conjugated to BD Horizon Red 718, which has been developed exclusively for BD Biosciences as a better alternative to Alexa Fluor® 700. BD Horizon Red 718 can be excited by the red laser (628 – 640 nm) and, with an Em Max around 718 nm, it can be detected using a 730/45 nm filter. Due to similar excitation and emission properties, we do not recommend using R718 in combination with APC-R700 or Alexa Fluor® 700.

566982 Rev. 3
Format Details
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R718
The BD Horizon™ Red 718 (R718) Dye is part of the BD red family of dyes. It is a small organic fluorochrome with an excitation maximum (Ex Max) at 695-nm and an emission maximum (Em Max) at 718-nm. Driven by BD innovation, R718 is designed to be excited by the red laser (627–640-nm) and detected using an optical filter centered near 720-nm (e.g., a 720/40-nm bandpass filter). R718 is a brighter alternative to Alexa Fluor™ 700. R718 is also a bright small molecule alternative to APC-R700 with lower spread into the APC detector. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
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R718
Red 627-640 nm
695 nm
718 nm
566982 Rev.3
Citations & References
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Development References (4)

  1. Chen H, Kinzer CA, Paul WE. p161, a murine membrane protein expressed on mast cells and some macrophages, is mouse CD13/aminopeptidase N. J Immunol. 1996; 157(6):2593-2600. (Clone-specific: Flow cytometry). View Reference
  2. Hansen AS, Noren O, Sjostrom H, Werdelin O. A mouse aminopeptidase N is a marker for antigen-presenting cells and appears to be co-expressed with major histocompatibility complex class II molecules. Eur J Immunol. 1993; 23(9):2358-2364. (Immunogen: Blocking, ELISA, Flow cytometry, Functional assay, Immunoprecipitation, Radioimmunoassay). View Reference
  3. Leenen PJ, Melis M, Kraal G, Hoogeveen AT, Van Ewijk W. The monoclonal antibody ER-BMDM1 recognizes a macrophage and dendritic cell differentiation antigen with aminopeptidase activity. Eur J Immunol. 1992; 22(6):1567-1572. (Biology). View Reference
  4. Pulendran B, Lingappa J, Kennedy MK, et al. Developmental pathways of dendritic cells in vivo: distinct function, phenotype, and localization of dendritic cell subsets in FLT3 ligand-treated mice. J Immunol. 1997; 159(5):2222-2231. (Biology). View Reference
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566982 Rev. 3

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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.