PE Rat Anti-Mouse C3aR

BD Pharmingen™ PE Rat Anti-Mouse C3aR

Name: PE Rat Anti-Mouse C3aR
Brand: BD Pharmingen™
SKU: 568276

Clone 14D4

(RUO)

Flow cytometric analysis of C3aR expression on mouse J774A.1 cells. Cells from the J774A.1 (Mouse macrophage, ATCC TIB-67) cell line were preincubated with Purified Rat Anti-Mouse CD16/CD32 antibody (Mouse BD Fc Block™) (Cat. No. 553141/553142) and then stained with either PE Rat IgG2a, κ Isotype Control (Cat No. 553930; dashed line histogram) or PE Rat Anti-Mouse C3aR antibody (Cat No. 568276: solid line histogram) at 0.5 µg/test. The fluorescence histogram showing C3aR expression (or Ig Isotype control staining) was derived from gated events with the forward and side light-scatter characteristics of viable mouse J774A.1 cells. Flow cytometric analysis was performed using a BD LSRFortessa™ Flow Cytometer System and FlowJo™ software. Data shown on this Technical Data Sheet are not lot specific.

Flow cytometric analysis of C3aR expression on mouse Peritoneal Exudate Cells (PECs). C57BL/6 mouse PECs were preincubated with Purified Rat Anti-Mouse CD16/CD32 antibody (Mouse BD Fc Block™) (Cat. No. 553141/553142). The cells were then stained with Alexa Fluor™ 647 Rat Anti-Mouse F4/80 antibody (Cat No. 565853) and with either PE Rat IgG2a, κ Isotype Control (Cat No. 553930; Left Plot) or PE Rat Anti-Mouse C3aR antibody (Cat No. 568276; Right Plot) at 0.5 µg/test. The two-color flow cytometric contour plot showing the corelated expression of C3aR (or Ig Isotype control staining) versus F4/80 was derived from gated events with the forward and side light-scatter characteristics of viable PECs. Flow cytometry and data analysis were performed using a BD FACSCanto™ Cell Analyzer System and FlowJo™ software. Data shown on this Technical Data Sheet are not lot specific.

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Product Details

Brand: BD Pharmingen™

Alternative Name: C3AR; C3a-R; C3ar1; anaphylatoxin C3a receptor

Reactivity: Mouse (QC Testing)

Isotype: Rat LOU, also known as Louvain, LOU/C, LOU/M IgG2a, κ

Immunogen: Mouse C3aR Transfected Cell Line

Application: Flow cytometry (Routinely Tested)

Concentration: 0.2 mg/ml

Storage Buffer: Aqueous buffered solution containing ≤0.09% sodium azide.

Regulatory Status: 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 unconjugated antibody and free dye were 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 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.
An isotype control should be used at the same concentration as the antibody of interest.
Please refer to http://regdocs.bd.com to access safety data sheets (SDS).

Companion Products

Stain Buffer (FBS) RUO

Size 500 mL Cat No. 554656

Stain Buffer (BSA) RUO

Size 500 mL Cat No. 554657

Purified Rat Anti-Mouse CD16/CD32 (Mouse BD Fc Block™) RUO

Size 0.5 mg Cat No. 553142

PE Rat IgG2a, κ Isotype Control RUO

Size 0.1 mg Cat No. 553930

Alexa Fluor® 647 Rat Anti-Mouse F4/80 RUO

Size 0.1 mg Cat No. 565853

568276 Rev. 1

Antibody Details

14D4

The 14D4 monoclonal antibody specifically recognizes the mouse C3a anaphylatoxin chemotactic receptor which is also known as C3aR. C3aR is a 54 kDa glycoprotein that is encoded by C3ar1 (Complement component 3a receptor 1) which belongs to the seven-transmembrane G-protein-coupled receptor (GPCR) superfamily. C3aR is variably expressed on monocytes, macrophages, dendritic cells, neutrophils, basophils, eosinophils, mast cells, T lymphocytes and B lymphocytes. C3aR functions as the cell surface receptor for the C3a anaphylatoxin, the C-terminal cleavage product of C3. C3a acts through C3aR to trigger granulocyte and mast cell degranulation that can further elicit histamine release, enhanced vascular permeability and smooth muscle contraction. C3a has chemotactic properties for eosinophils and mast cells. C3a also causes release of key cytokines from multiple cell types, including IL-1β, IL-6, and TNF.

568276 Rev. 1

Format Details

R-Phycoerythrin (PE), is part of the BD family of Phycobiliprotein dyes. This fluorochrome is a multimeric fluorescent phycobiliprotein with excitation maximum (Ex Max) of 496 nm and 566 nm and an emission maximum (Em Max) at 576 nm. PE is designed to be excited by the Blue (488 nm), Green (532 nm) and Yellow-Green (561 nm) lasers and detected using an optical filter centered near 575 nm (e.g., a 575/26-nm bandpass filter). As PE is excited by multiple lasers, this can result in cross-laser excitation and fluorescence spillover on instruments with various combinations of Blue, Green, and Yellow-Green lasers. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.

Format: PE

Excitation Source: Yellow-Green 488 nm, 532 nm, 561 nm

Excitation Max: 496 nm, 566 nm

Emission Max: 576 nm

568276 Rev.1

Citations & References

View product citations for antibody "568276" on CiteAb

Development References (3)

Guglietta S, Chiavelli A, Zagato E, et al. Coagulation induced by C3aR-dependent NETosis drives protumorigenic neutrophils during small intestinal tumorigenesis.. Nat Commun. 2016; 7:11037. (Clone-specific: Flow cytometry). View Reference
Kiafard Z, Tschernig T, Schweyer S, Bley A, Neumann D, Zwirner J. Use of monoclonal antibodies to assess expression of anaphylatoxin receptors in tubular epithelial cells of human, murine and rat kidneys.. Immunobiology. 2007; 212(2):129-39. (Immunogen: Flow cytometry, Immunohistochemistry). View Reference
Quell KM, Karsten CM, Kordowski A, et al. Monitoring C3aR Expression Using a Floxed tdTomato-C3aR Reporter Knock-in Mouse.. J Immunol. 2017; 199(2):688-706. (Clone-specific: Flow cytometry). View Reference

568276 Rev. 1

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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.