Multicolor flow cytometric analysis of CD146 expression on C57BL/6 splenocytes. Spleen cells were preincubated with Mouse BD Fc Block™ purified anti-mouse CD16/CD32 mAb 2.4G2 (Cat. No. 553141/553142) and then stained with PE Rat Anti-Mouse CD49b antibody (Cat. No. 561066, Left and Middle Panel) or APC Mouse Anti-Mouse NK-1.1 antibody (Cat. No. 550627/561117. Right Panel) and with either Alexa Fluor® 488 Rat IgG2a, κ Isotype Control (Cat. No. 557676; Left Panel) or Alexa Fluor® 488 Rat Anti-Mouse CD146 antibody (Cat. No. 562229; Middle and Right Panel). Two-color flow cytometric dot plots showing the correlated expression patterns of CD49b or NK-1.1 versus CD146 (or Ig isotype control staining) were derived from gated events with the forward and side light-scatter characteristics of viable splenocytes. Flow cytometry was performed using a BD™ LSR II Flow Cytometer System.
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Multicolor flow cytometric analysis of CD146 expression on C57BL/6 splenocytes. Spleen cells were preincubated with Mouse BD Fc Block™ purified anti-mouse CD16/CD32 mAb 2.4G2 (Cat. No. 553141/553142) and then stained with PE Rat Anti-Mouse CD49b antibody (Cat. No. 561066, Left and Middle Panel) or APC Mouse Anti-Mouse NK-1.1 antibody (Cat. No. 550627/561117. Right Panel) and with either Alexa Fluor® 488 Rat IgG2a, κ Isotype Control (Cat. No. 557676; Left Panel) or Alexa Fluor® 488 Rat Anti-Mouse CD146 antibody (Cat. No. 562229; Middle and Right Panel). Two-color flow cytometric dot plots showing the correlated expression patterns of CD49b or NK-1.1 versus CD146 (or Ig isotype control staining) were derived from gated events with the forward and side light-scatter characteristics of viable splenocytes. Flow cytometry was performed using a BD™ LSR II Flow Cytometer System.
Product Details
BD Pharmingen™
S-endo 1; S-endo; Muc18; Mcam; Gicerin
Mouse (QC Testing)
Rat IgG2a, κ
CBA/J Mouse lymph node Cell Line
Flow cytometry (Routinely Tested)
0.2 mg/ml
84004
AB_11153320
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 Alexa Fluor® 488 under optimum conditions, and unreacted Alexa Fluor® 488 was removed.
Product Notices
- Since applications vary, each investigator should titrate the reagent to obtain optimal results.
- An isotype control should be used at the same concentration as the antibody of interest.
- 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® 488 fluorochrome emission is collected at the same instrument settings as for fluorescein isothiocyanate (FITC).
- 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.
- 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.
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
Alexa Fluor® 488 Rat IgG2a, κ Isotype Control RUO
Size
0.1 mg
Cat No.
557676
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PE Rat Anti-Mouse CD49b RUO
Size
25 µg
Cat No.
561066
APC Mouse Anti-Mouse NK-1.1 RUO
Size
0.1 mg
Cat No.
550627
562229 Rev. 1
Antibody Details
ME-9F1
The ME-9F1 monoclonal antibody specifically binds to mouse CD146. The CD146 adhesion molecule is a type 1 transmembrane glycoprotein and member of the immunoglobulin superfamily. CD146 is expressed by blood vessel endothelial cells and may play roles in forming intercellular junctions between endothelial cells and influencing the transendothelial migration of other cell types. CD146 is also expressed by some melanoma cell lines, NK cells and neutrophils. CD146 is not detectable on mouse monocytes, dendritic cells, T cells, NKT cells, B cells and smooth muscle cells. Increased expression of CD146 is reportedly associated with NK cell maturation and may be used to characterize different functional NK cell subsets. Activated CD146-positive mouse NK cells reportedly are less cytotoxic and secrete less IFN-γ than their CD146-negative counterparts.
562229 Rev. 1
Format Details
Alexa Fluor™ 488
Alexa Fluor™ 488 Dye is part of the BD blue family of dyes. This is a small organic fluorochrome with an excitation maximum (Ex Max) at 494-nm and an emission maximum (Em Max) at 517-nm. Alexa Fluor™ 488 is designed to be excited by the Blue laser (488 nm) and detected using an optical filter centered near 520-nm (e.g., a 530/30-nm bandpass filter). Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
Alexa Fluor™ 488
Blue 488 nm
494 nm
517 nm
562229 Rev.1
Citations & References
Development References (4)
-
Alais S, Allioli N, Pujades C, et al. HEMCAM/CD146 downregulates cell surface expression of beta1 integrins. J Cell Sci. 2001; 114(Pt 10):1847-1859. (Biology). View Reference
-
Despoix N, Walzer T, Jouve N, et al. Mouse CD146/MCAM is a marker of natural killer cell maturation. Eur J Immunol. 2008; 38(10):2855-2864. (Biology). View Reference
-
Schrage A, Loddenkemper C, Erben U, et al. Murine CD146 is widely expressed on endothelial cells and is recognized by the monoclonal antibody ME-9F1. Histochem Cell Biol. 2008; 129(4):441-451. (Clone-specific: Flow cytometry, Immunofluorescence, Immunohistochemistry, Immunoprecipitation). View Reference
-
Yang H, Wang S, Liu Z, et al. Isolation and characterization of mouse MUC18 cDNA gene, and correlation of MUC18 expression in mouse melanoma cell lines with metastatic ability. Gene. 2001; 265(1-2):133-145. (Biology). View Reference
562229 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.
