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Methanol-fixed MCF-7 cells were stained with CD63 Mouse Monoclonal Antibody (H5C6) followed by goat anti-mouse CF® 568. The nuclei were stained with DAPI, and the cells were imaged in the Cy® 3 and DAPI channels on a Zeiss LSM 700 confocal microscope using a 40X oil objective. Western detection of human CD63 in MCF-7 cell lysates using CD63 Mouse Monoclonal Antibody (H5C6) followed by goat anti-mouse CF® 680. M: 1.5 uL Peacock™ Prestained Protein Marker. Lane 1: 10 ug MCF-7 cell lysate. Lane 2: 1 ug MCF-7 cell lysate. The blots were imaged on a LI-COR Odyssey® infrared imaging system in the 700 channel. Flow cytometry of fixed and permeabilized U2-OS cells stained with CD63 Mouse Monoclonal Antibody (H5C6), CF® 405M Conjugate (blue) or with an IgG1 isotype control antibody, CF® 405M Conjugate (gray). The staining was detected in the V450 (Pacific Blue) channel. Flow cytometry of U2-OS cells stained with CD63 Mouse Monoclonal Antibody (H5C6), CF® 488A Conjugate (green) or with an IgG1 isotype control antibody, CF® 488A Conjugate (gray). The staining was detected in the B525 (FITC) channel. Flow cytometry of MCF-7 cells unstained (gray) or stained with CD63 Mouse Monoclonal Antibody (H5C6), R-PE Conjugate (orange). Staining was detected in the Y585 (PE) channel. Live MCF-7 cells were labeled with CD63 Mouse Monoclonal Antibody (H5C6) followed by goat anti-mouse CF® 568 (orange histogram). Cells stained with an IgG1 isotype control antibody and the same secondary show no staining (gray histogram). Staining was detected in the Y585 (PE) channel.
Flow cytometry of U2-OS cells stained with CD63 Mouse Monoclonal Antibody (H5C6), CF® 568 Conjugate (orange) or with an IgG1 isotype control antibody, CF® 568 Conjugate (gray). The staining was detected in the Y585 (PE) channel. Flow cytometry of MCF-7 cells stained with CD63 Mouse Monoclonal Antibody (H5C6), CF® 700 conjugate (teal) or with an IgG1 isotype control antibody, CF® 700 conjugate (gray). The staining was detected in the R712 (APC-700) channel.
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Product Description
Note: CD63 clone H5C6 is being discontinued and has been replaced by CD63 Recombinant Monoclonal Mouse Antibody (2304.63) – Biotium Choice, which has been validated to have the same bright and specific staining of human CD63.
CD63 (clone H5C6) is a mouse monoclonal antibody that recognizes the human tetraspanin protein CD63. This antibody belongs to the Biotium Choice list of select antibodies that have been validated and optimized in-house for optimal performance. This antibody has been validated in flow cytometry, in immunofluorescence microscopy, and in western blot. The antibody is available conjugated to a selection of bright CF® Dyes. It is supplied in PBS, 0.1% BSA, 0.05% azide or purified at 0.5 mg/mL, BSA-free.
The tetraspanins are integral membrane proteins expressed on cell surface and granular membranes of hematopoietic cells and are components of multi-molecular complexes with specific integrins. The tetraspanin CD63 is a lysosomal membrane glycoprotein that translocates to the plasma membrane after platelet activation. CD63 is expressed on activated platelets, monocytes and macrophages, and is weakly expressed on granulocytes, T cell and B cells. It is located on the basophilic granule membranes and on the plasma membranes of lymphocytes and granulocytes. CD63 is a member of the TM4 superfamily of leukocyte glycoproteins that includes CD9, CD37 and CD53, which contain four transmembrane regions. CD63 may play a role in phagocytic and intracellular lysosome-phagosome fusion events. CD63 deficiency is associated with Hermansky-Pudlak syndrome and is strongly expressed during the early stages of melanoma progression.
Tetraspanin proteins CD9, CD63, and CD81, are the most common proteins used as exosome markers. Tetraspanins are plasma membrane proteins with many proposed functions, including activation and sorting of other membrane proteins. They are also thought to play a role in the targeting of proteins to multivesicular bodies (MVBs) and exosomes. These tetraspanins are broadly expressed on many cell types and can therefore be detected on many types of exosomes, but their expression levels vary depending on the cell type of origin.
As an isotype control, we recommend Isotype Control, Monoclonal Mouse IgG1 Kappa (IGG1/1331) for this antibody. View our full catalog of Biotium Choice antibodies, optimized and validated for the best results in flow cytometry.
Exosome Stains and Antibodies
Biotium also offers ExoBrite™ CD63 Flow Antibody and ExoBrite™ CD63 Western Antibody that are validated for optimal detection of isolated exosomes by flow cytometry or western blotting. Learn more about our products for exosome research.
Discover Sharper Signals and Unmatched Panel Flexibility with Biotium Choice Antibodies Powered by CF® Dyes and Astral Leap™ Tandem Dyes Biotium Choice antibodies are carefully curated and validated in-house to offer exceptional signal-to-noise. Labeled with our advanced CF® Dyes and Astral Leap™ tandem dyes, they are our top-recommended antibodies for flow cytometry and other applications.
Biotium Choice Antibody Features
Robust and validated clones against common targets
Developed and optimized for flow cytometry and other applications
Conjugated to bright, photostable CF® Dyes for superior signal and clarity
Also available with Astral Leap™ tandem dyes for expanding multiplexing
New antibody clones and dye conjugates continuously in development
View our full catalog of Biotium Choice antibodies
Identifying Biotium validated antibodies
Biotium scientists validate and optimize select antibodies in our catalog, marked with a check mark next to the product name. This symbol indicates the antibody is highly recommended for the specified application(s).
Look for this symbol next to the product name on product pages and search results
CD63 Monoclonal Mouse Antibody (H5C6) - Biotium Choice
Conjugation Ex/Em Conc. Size Catalog No. Availability
CF® 405M 416/452 nm 100 ug/mL 25 tests (125 uL) P004S-405M-125 Discontinued, see replacement P013-405M
100 tests (500 uL) P004S-405M-500
CF® 488A 490/516 nm 100 ug/mL 25 tests (125 uL) P004S-488A-125 Discontinued, see replacement P013-488A
100 tests (500 uL) P004S-488A-500
CF® 568 562/584 nm 100 ug/mL 25 tests (125 uL) P004S-568-125 Discontinued, see replacement P013-568
100 tests (500 uL) P004S-568-500
CF® 700 696/721 nm 100 ug/mL 25 tests (125 uL) P004S-700-125 Discontinued, see replacement P013-700
100 tests (500 uL) P004S-700-500
R-PE 496, 546, 565/576 nm 100 ug/mL 25 tests (125 uL) P004S-RPE-125 Discontinued, see replacement P013-RPE
100 tests (500 uL) P004S-RPE-500
Purified (BSA-free) N/A 0.5 mg/mL 200 uL (100 ug) P004-200uL Available
Technology
Primary Antibody Conjugates
Western Blotting & Protein Analysis
Tech Tips
Troubleshooting Tips for Fluorescence Staining
Protocol: Immunofluorescence Staining of Cells for Microscopy
Tech Tip: Avoiding Artifacts from UV Photoconversion of DAPI and Hoechst
Tech Tip: Fluorescence Bleed-Through and Cross-Talk
Product Attributes
Biotium Choice Antibody
Primary
Antibody type
Biotium Choice, Primary
Antibody reactivity (target)
CD63
Synonyms
gp55; granulophysin; Lysosomal-associated membrane protein 3 (LAMP-3); Mast cell antigen AD1; melanoma 1 antigen; Melanoma-associated antigen MLA1; Melanoma-associated antigen ME491; MLA1; NGA; Ocular melanoma-associated antigen; OMA81H; PTLGP40; Tetraspanin-30; TSPAN30
Species reactivity
Baboon, Cynomolgus monkey, Human, Non-human primates
Molecular weight
26 kDa (core protein); 30-60 kDa (glycosylated)
Antibody target cellular localization
Exosomes/EVs, Lysosomes, Plasma membrane, Membrane/vesicular, Multivesicular bodies
Cell/tissue expression
Exosomes, Platelets, Granulocytes, Lymphocytes, Monocytes/macrophages
Verified antibody applications
Flow (surface) (verified), IF (verified), WB (verified)
Positive control
U87MG, SK-MEL-28, HL60, THP-1, NIH/3T3, or MCF-7 cells. Human melanoma, spleen or lymphoma tissue.
Antibody application notes
Flow Cytometry: 0.5-1 ug/mL (or, 0.05-0.1 ug/million cells), Immunofluorescence: 1 ug/mL, Western Blot: 0.1 ug/mL
Antibody research areas
Exosomes/EVs
Antibody/conjugate formulation
Purified: 0.5 mg/mL in PBS/0.05% azide, Conjugates: PBS/0.1% BSA/0.05% azide
Shelf life
Guaranteed for at least 24 months from date of receipt when stored as recommended
Volume per assay
5 uL/test
Storage Conditions
Store at 2 to 8 °C
Regulatory status
For research use only (RUO)
Product origin
Product may contain either bovine serum albumin (BSA) from bovine serum (Bos taurus), or recombinant BSA produced in Chinese hamster ovary cells. Inquire for the specific lot.
Documents, Protocols, SDS and COA
Datasheet
Datasheet Download
COA Request the Certificate of Analysis
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