Two-color flow cytometric analysis of S100A9 (MRP-14) expression in mouse bone marrow cells. C57BL/6 mouse bone marrow cells 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 Ly-6G and LY-6C antibody (Cat. No. 553128/561084). The cells were washed, and then fixed and permeabilized using the BD Cytofix/Cytoperm™ Fixation/Permeablization Kit (Cat. No. 554714) and stained with either Alexa Fluor® 647 Rat IgG1, κ Isotype Control (Cat. No. 557731; Left Plot) or Alexa Fluor® 647 Rat Anti-Mouse S100A9 (MRP-14) antibody (Cat. No. 565833; Right Plot). Two-color flow cytometric contour plots showing the correlated expression of S100A9 (MRP-14) [or Ig Isotype control staining] versus Ly-6G and LY-6C were derived from gated events with the forward and side light-scatter characteristics of intact bone marrow leucocytes. Flow cytometric analysis was performed using a BD LSRFortessa™ X-20 Cell Analyzer System.
Two-color flow cytometric analysis of S100A9 (MRP-14) expression in mouse bone marrow cells. C57BL/6 mouse bone marrow cells 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 Ly-6G and LY-6C antibody (Cat. No. 553128/561084). The cells were washed, and then fixed and permeabilized using the BD Cytofix/Cytoperm™ Fixation/Permeablization Kit (Cat. No. 554714) and stained with either Alexa Fluor® 647 Rat IgG1, κ Isotype Control (Cat. No. 557731; Left Plot) or Alexa Fluor® 647 Rat Anti-Mouse S100A9 (MRP-14) antibody (Cat. No. 565833; Right Plot). Two-color flow cytometric contour plots showing the correlated expression of S100A9 (MRP-14) [or Ig Isotype control staining] versus Ly-6G and LY-6C were derived from gated events with the forward and side light-scatter characteristics of intact bone marrow leucocytes. Flow cytometric analysis was performed using a BD LSRFortessa™ X-20 Cell Analyzer System.
Product Details
BD Pharmingen™
S100a9; MRP-14; Calgranulin-B; GAGB; Cagb; BEE22; L1Ag; 60B8Ag; AW546964
Mouse (QC Testing)
Rat IgG1, κ
Mouse S100A9 Recombinant Protein
Intracellular staining (flow cytometry) (Routinely Tested)
0.2 mg/ml
AB_2739373
Aqueous buffered solution containing ≤0.09% sodium azide.
RUO
Preparation And Storage
The antibody was conjugated to Alexa Fluor® 647 under optimum conditions, and unreacted Alexa Fluor® 647 was removed. 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.
Product Notices
- Since applications vary, each investigator should titrate the reagent to obtain optimal results.
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
- 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® 647 fluorochrome emission is collected at the same instrument settings as for allophycocyanin (APC).
- Alexa Fluor® is a registered trademark of Molecular Probes, Inc., Eugene, OR.
- 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.
- An isotype control should be used at the same concentration as the antibody of interest.
Companion Products
Purified Rat Anti-Mouse CD16/CD32 (Mouse BD Fc Block™) RUO
Size
0.1 mg
Cat No.
553141
Purified Rat Anti-Mouse CD16/CD32 (Mouse BD Fc Block™) RUO
Size
0.5 mg
Cat No.
553142
Stain Buffer (FBS) RUO
Size
500 mL
Cat No.
554656
Stain Buffer (BSA) RUO
Size
500 mL
Cat No.
554657
Fixation/Permeabilization Kit RUO
Size
250 Tests
Cat No.
554714
Alexa Fluor® 647 Rat IgG1, κ Isotype Control RUO
Size
0.1 mg
Cat No.
557731
565833 Rev. 1
Antibody Details
2B10
The 2B10 monoclonal antibody specifically binds to S100 calcium-binding protein A9 (S100A9), which is also known as Migration inhibitory factor-related protein 14 (MRP-14) or Calgranulin-B (CAGB). S100A9 is a member of the S100 family of calcium and zinc binding proteins. S100A9 is predominantly expressed in myeloid cells, such as, neutrophils and monocytes. It is one of the most abundant proteins expressed in myeloid cells. Similar to other S100 family of proteins, S100A9 functions as a calcium sensor and may enhance the chemotaxis and proinflammatory response of monocytes and neutrophils.
565833 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
565833 Rev.1
Citations & References
View product citations for antibody "565833" on CiteAb
Development References (4)
-
Baker JR1, Jeffery R, May RD, et al. Dev Dyn. 2011; 240(9):2194-2203. (Clone-specific: Western blot). View Reference
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Foell D, Wittkowski H, Vogl T, Roth J. S100 proteins expressed in phagocytes: a novel group of damage-associated molecular pattern molecules.. J Leukoc Biol. 2007; 81(1):28-37. (Biology). View Reference
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Hobbs JA, May R, Tanousis K, et al. Myeloid cell function in MRP-14 (S100A9) null mice.. Mol Cell Biol. 2003; 23(7):2564-76. (Immunogen: Flow cytometry, Immunohistochemistry, Western blot). View Reference
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Nacken W, Roth J, Sorg C, Kerkhoff C. S100A9/S100A8: Myeloid representatives of the S100 protein family as prominent players in innate immunity.. Microsc Res Tech. 2003; 60(6):569-80. (Biology). View Reference
565833 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.