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BD Pharmingen™ Purified Rat Anti-Mouse CD63
Clone NVG-2 (RUO)
Analyses of CD63 expression.
Panel 1 and 2. Multicolor flow cytometric analysis of CD63 expression on mouse bone marrow cells. C57BL/6 mouse bone marrow cells were stained with either Purified Rat IgG2a, κ Isotype Control (Cat. No. 553927; Panel 1) or Purified Rat Anti-Mouse CD63 antibody (Cat. No. 564221; Panel 2). The cells were washed and counterstained with PE Goat Anti-Rat Ig (Cat. No. 550767). After washing, the cells were then stained with APC Rat Anti-Mouse Ly-6G and Ly-6C antibody (Cat. No. 553129/561083). Two-color flow cytometric dot plots show the correlated expression patterns of Ly-6G and Ly-6C versus CD63 (or Ig Isotype control staining) for gated events with the forward and side light-scatter characteristics of viable bone marrow cells. Flow cytometric analysis was performed using a BD LSRFortessa™ Cell Analyzer System.
Panel 3. Immunofluorescent staining of CD63 in bEnd.3 cells. Adherent bEnd.3 cells (ATCC CRL-2299) were fixed with BD Cytofix™ Fixation Buffer (Cat. No. 554655), permeabilized with 0.1% Triton™ X-100 (Sigma, X100), and stained with either Purified Rat IgG2a, κ Isotype Control (Upper Panel 3) or Purified Rat Anti-Mouse CD63 antibody (Lower Panel 3). The cells were washed and then counterstained with Alexa Fluor® 488 Goat Anti-Rat IgG (H+L) Antibody (Life Technologies, Cat. No. A11006; pseudocolored green) and Hoechst 33342 Solution (Cat. No. 561908; pseudocolored blue). Images were analyzed using a BD Pathway™ 435 cell imager and merged using BD AttoVision™ Software.
Analyses of CD63 expression.
Panel 1 and 2. Multicolor flow cytometric analysis of CD63 expression on mouse bone marrow cells. C57BL/6 mouse bone marrow cells were stained with either Purified Rat IgG2a, κ Isotype Control (Cat. No. 553927; Panel 1) or Purified Rat Anti-Mouse CD63 antibody (Cat. No. 564221; Panel 2). The cells were washed and counterstained with PE Goat Anti-Rat Ig (Cat. No. 550767). After washing, the cells were then stained with APC Rat Anti-Mouse Ly-6G and Ly-6C antibody (Cat. No. 553129/561083). Two-color flow cytometric dot plots show the correlated expression patterns of Ly-6G and Ly-6C versus CD63 (or Ig Isotype control staining) for gated events with the forward and side light-scatter characteristics of viable bone marrow cells. Flow cytometric analysis was performed using a BD LSRFortessa™ Cell Analyzer System.
Panel 3. Immunofluorescent staining of CD63 in bEnd.3 cells. Adherent bEnd.3 cells (ATCC CRL-2299) were fixed with BD Cytofix™ Fixation Buffer (Cat. No. 554655), permeabilized with 0.1% Triton™ X-100 (Sigma, X100), and stained with either Purified Rat IgG2a, κ Isotype Control (Upper Panel 3) or Purified Rat Anti-Mouse CD63 antibody (Lower Panel 3). The cells were washed and then counterstained with Alexa Fluor® 488 Goat Anti-Rat IgG (H+L) Antibody (Life Technologies, Cat. No. A11006; pseudocolored green) and Hoechst 33342 Solution (Cat. No. 561908; pseudocolored blue). Images were analyzed using a BD Pathway™ 435 cell imager and merged using BD AttoVision™ Software.
Product Details
BD Pharmingen™
Cd63; Tspan30; Melanoma 1 antigen; ME491; C75951
Mouse (QC Testing)
Rat SD, also known as Sprague-Dawley (outbred) IgG2a, κ
Mouse Eosinophils
Flow cytometry (Routinely Tested), Bioimaging, Immunofluorescence, Intracellular staining (flow cytometry) (Tested During Development)
0.5 mg/ml
AB_2738677
Aqueous buffered solution containing ≤0.09% sodium azide.
RUO
Preparation And Storage
Store undiluted at 4°C. 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.
- 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.
- Sodium azide is a reversible inhibitor of oxidative metabolism; therefore, antibody preparations containing this preservative agent must not be used in cell cultures nor injected into animals. Sodium azide may be removed by washing stained cells or plate-bound antibody or dialyzing soluble antibody in sodium azide-free buffer. Since endotoxin may also affect the results of functional studies, we recommend the NA/LE (No Azide/Low Endotoxin) antibody format, if available, for in vitro and in vivo use.
- 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.
- Alexa Fluor® is a registered trademark of Molecular Probes, Inc., Eugene, OR.
- Triton is a trademark of the Dow Chemical Company.
Companion Products
Stain Buffer (FBS) RUO
Size
500 ML
Cat No.
554656
Stain Buffer (BSA) RUO
Size
500 ML
Cat No.
554657
Purified Rat IgG2a, κ Isotype Control RUO
Size
0.5 mg
Cat No.
553927
PE Goat Anti-Rat Ig RUO
Size
0.2 mg
Cat No.
550767
Hoechst 33342 Solution RUO
Size
1 mg
Cat No.
561908
564221 Rev. 1
Antibody Details
NVG-2
The NVG-2 monoclonal antibody specifically binds to CD63 which is also known as Tspan30, Melanoma 1 antigen, ME491, and C75951. CD63 is a tetraspanin which belongs to the transmembrane-4 glycoprotein superfamily (TM4SF). It constitutes a major component of lysosomal membranes and translocates to the cell surface in response to various stimuli. CD63 is expressed by activated platelets, granulocytes, monocytes, macrophages, endothelium, fibroblasts, osteoblasts, and smooth muscle cells. CD63 can associate with other membrane molecules and functions in cellular signaling, adhesion and motility.
564221 Rev. 1
Format Details
Purified
Tissue culture supernatant is purified by either protein A/G or affinity purification methods. Both methods yield antibody in solution that is free of most other soluble proteins, lipids, etc. This format provides pure antibody that is suitable for a number of downstream applications including: secondary labeling for flow cytometry or microscopy, ELISA, Western blot, etc.
Purified
564221 Rev.1
Citations & References
View product citations for antibody "564221" on CiteAb
Development References (2)
-
Miyamoto H, Homma M, Hotta H. Molecular cloning of the murine homologue of CD63/ME491 and detection of its strong expression in the kidney and activated macrophages. Biochim Biophys Acta. 1994; 1217(3):312-316. (Biology). View Reference
-
Verjan Garcia N, Umemoto E, Saito Y, et al. SIRPalpha/CD172a regulates eosinophil homeostasis. J Immunol. 2011; 187(5):2268-2277. (Immunogen: Flow cytometry, Fluorescence microscopy, Immunofluorescence). View Reference
564221 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.