Key Product Details

Species Reactivity

Validated:

Human

Cited:

Human, Porcine

Applications

Validated:

Immunohistochemistry, Western Blot, Flow Cytometry, Simple Western, CyTOF-ready

Cited:

Western Blot, Simple Western

Label

Unconjugated

Antibody Source

Recombinant Monoclonal Rabbit IgG Clone # 2585J
View all CD63 Primary Antibodies »

Product Specifications

Immunogen

Chinese Hamster Ovary cell line CHO-derived human CD63
Ala103-Val203
Accession # P08962-1

Specificity

Detects human CD63 in direct ELISAs.

Clonality

Monoclonal

Host

Rabbit

Isotype

IgG

Scientific Data Images for Human CD63 Antibody

Detection of Human CD63 antibody by Western Blot.

Detection of Human CD63 by Western Blot.

Western blot shows lysates of human platelets. PVDF membrane was probed with 0.5 µg/mL of Rabbit Anti-Human CD63 Monoclonal Antibody (Catalog # MAB50482) followed by HRP-conjugated Anti-Rabbit IgG Secondary Antibody (HAF008). A specific band was detected for CD63 at approximately 55 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

CD63 antibody in Human Lung Cancer Tissue by Immunohistochemistry (IHC-P).

CD63 in Human Lung Cancer Tissue.

CD63 was detected in immersion fixed paraffin-embedded sections of human lung cancer tissue using Rabbit Anti-Human CD63 Monoclonal Antibody (Catalog # MAB50482) at 0.3 µg/mL for 1 hour at room temperature followed by incubation with the Anti-Rabbit IgG VisUCyte™ HRP Polymer Antibody (VC003). Before incubation with the primary antibody, tissue was subjected to heat-induced epitope retrieval using Antigen Retrieval Reagent-Basic (Catalog # CTS013). Tissue was stained using DAB (brown) and counterstained with hematoxylin (blue). Specific staining was localized to cell membrane in sperm cells. View our protocol for IHC Staining with VisUCyte HRP Polymer Detection Reagents.

Detection of CD63 antibody in Human PBMC antibody by Flow Cytometry.

Detection of CD63 in Human PBMC by Flow Cytometry.

Human PBMC were stained with Rabbit Anti-Human CD63 Monoclonal Antibody (Catalog # MAB50482, filled histogram) or isotype control antibody (Catalog # MAB1050, open histogram), followed by Phycoerythrin-conjugated Anti-Rabbit IgG Secondary Antibody (F0111). To facilitate intracellular staining, cells were fixed and permeabilized with FlowX FoxP3/Transcription Factor Fixation & Perm Buffer Kit (FC012). View our protocol for Staining Intracellular Molecules.

Detection of Human CD63 antibody by Simple WesternTM.

Detection of Human CD63 by Simple WesternTM.

Simple Western lane view shows lysates of human platelets, loaded at 0.2 mg/mL. A specific band was detected for CD63 at approximately 59 kDa (as indicated) using 20 µg/mL of Rabbit Anti-Human CD63 Monoclonal Antibody (Catalog # MAB50482). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.

Detection of CD63 in PBMC monocytes by Flow Cytometry

PBMC monocytes were stained with Rabbit Anti-Human CD63 Monoclonal Antibody (Catalog # MAB50482, filled histogram) or isotype control antibody (Catalog # MAB1050, open histogram) followed by Phycoerythrin-conjugated Anti-Rabbit IgG Secondary Antibody (Catalog # F0110). View our protocol for Staining Membrane-associated Proteins.

Detection of CD63 by Western Blot

Detection of CD63 by Western Blot

Purified exosome product (PEP) characterization, quantification, and delivery biodistribution. A NanoSight nanoparticle analysis of size distribution and concentration of PEP diluted in phosphate buffered saline 1:1000 documented 6.65 × 1012 ± 1.16 × 1011. B Transmission electron microscopy of PEP. Scale = 200 nm arrow heads pointing to EVs. C Western blot probing for CD63, CD9, and Flotillin-1 in 3 separate CGMP manufactured PEP lots. D Western blot comparison of NF-kappa B p65 and PD-L1 levels, in 3 separate CGMP manufactured PEP lots versus adipose-derived mesenchymal stem cell conditioned media (AMSC-CM). E Atomic-force microscope comparing platelet-conditioned medium EV isolation using centrifugation versus the PEP process, scale bar embedded in the image. F Representative image from Single-particle interferometric reflectance imaging sensing (SP-IRIS) analysis for presence of surface CD41a, CD9, CD63, and CD81 tetraspanins. G Graphical representation of the SP-IRIS analysis. H Quantitation of CD9, CD63 and CD81 on a CD41a captured plate documented vast majority of PEP as CD41a/CD9 positive, with smaller representation from CD63 and background CD81. Data presented as mean ± stdev. N = 3 separate CGMP manufactured PEP lots. I Pie chart representation of the exosome tetraspanin surface marker profile of CD41a captured PEP exosomes. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/36175423), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of CD63 by Western Blot

Detection of CD63 by Western Blot

Purified exosome product (PEP) characterization, quantification, and delivery biodistribution. A NanoSight nanoparticle analysis of size distribution and concentration of PEP diluted in phosphate buffered saline 1:1000 documented 6.65 × 1012 ± 1.16 × 1011. B Transmission electron microscopy of PEP. Scale = 200 nm arrow heads pointing to EVs. C Western blot probing for CD63, CD9, and Flotillin-1 in 3 separate CGMP manufactured PEP lots. D Western blot comparison of NF-kappa B p65 and PD-L1 levels, in 3 separate CGMP manufactured PEP lots versus adipose-derived mesenchymal stem cell conditioned media (AMSC-CM). E Atomic-force microscope comparing platelet-conditioned medium EV isolation using centrifugation versus the PEP process, scale bar embedded in the image. F Representative image from Single-particle interferometric reflectance imaging sensing (SP-IRIS) analysis for presence of surface CD41a, CD9, CD63, and CD81 tetraspanins. G Graphical representation of the SP-IRIS analysis. H Quantitation of CD9, CD63 and CD81 on a CD41a captured plate documented vast majority of PEP as CD41a/CD9 positive, with smaller representation from CD63 and background CD81. Data presented as mean ± stdev. N = 3 separate CGMP manufactured PEP lots. I Pie chart representation of the exosome tetraspanin surface marker profile of CD41a captured PEP exosomes. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/36175423), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of CD63 by Western Blot

Detection of CD63 by Western Blot

Purified exosome product (PEP) characterization, quantification, and delivery biodistribution. A NanoSight nanoparticle analysis of size distribution and concentration of PEP diluted in phosphate buffered saline 1:1000 documented 6.65 × 1012 ± 1.16 × 1011. B Transmission electron microscopy of PEP. Scale = 200 nm arrow heads pointing to EVs. C Western blot probing for CD63, CD9, and Flotillin-1 in 3 separate CGMP manufactured PEP lots. D Western blot comparison of NF-kappa B p65 and PD-L1 levels, in 3 separate CGMP manufactured PEP lots versus adipose-derived mesenchymal stem cell conditioned media (AMSC-CM). E Atomic-force microscope comparing platelet-conditioned medium EV isolation using centrifugation versus the PEP process, scale bar embedded in the image. F Representative image from Single-particle interferometric reflectance imaging sensing (SP-IRIS) analysis for presence of surface CD41a, CD9, CD63, and CD81 tetraspanins. G Graphical representation of the SP-IRIS analysis. H Quantitation of CD9, CD63 and CD81 on a CD41a captured plate documented vast majority of PEP as CD41a/CD9 positive, with smaller representation from CD63 and background CD81. Data presented as mean ± stdev. N = 3 separate CGMP manufactured PEP lots. I Pie chart representation of the exosome tetraspanin surface marker profile of CD41a captured PEP exosomes. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/36175423), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of CD63 by Western Blot

Detection of CD63 by Western Blot

Purified exosome product (PEP) characterization, quantification, and delivery biodistribution. A NanoSight nanoparticle analysis of size distribution and concentration of PEP diluted in phosphate buffered saline 1:1000 documented 6.65 × 1012 ± 1.16 × 1011. B Transmission electron microscopy of PEP. Scale = 200 nm arrow heads pointing to EVs. C Western blot probing for CD63, CD9, and Flotillin-1 in 3 separate CGMP manufactured PEP lots. D Western blot comparison of NF-kappa B p65 and PD-L1 levels, in 3 separate CGMP manufactured PEP lots versus adipose-derived mesenchymal stem cell conditioned media (AMSC-CM). E Atomic-force microscope comparing platelet-conditioned medium EV isolation using centrifugation versus the PEP process, scale bar embedded in the image. F Representative image from Single-particle interferometric reflectance imaging sensing (SP-IRIS) analysis for presence of surface CD41a, CD9, CD63, and CD81 tetraspanins. G Graphical representation of the SP-IRIS analysis. H Quantitation of CD9, CD63 and CD81 on a CD41a captured plate documented vast majority of PEP as CD41a/CD9 positive, with smaller representation from CD63 and background CD81. Data presented as mean ± stdev. N = 3 separate CGMP manufactured PEP lots. I Pie chart representation of the exosome tetraspanin surface marker profile of CD41a captured PEP exosomes. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/36175423), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of CD63 by Western Blot

Detection of CD63 by Western Blot

Production and characterization of engineered BMP2- and BMP7-EV.A Schematic of lentiviral vector with BMP2 (top) or BMP2 and BMP7 (bottom) joined with T2A linker. Acronyms are as follows: LTR (Long Terminal Repeat), Ψ (viral packaging), RRE (Rev response element), cPPT (central polypurine tract), WPRE (post-regulatory element), delta U3 (U3 deletion in 3’ LTR). B Real-time quantitative reverse-transcriptase PCR to show delta CT ratio to GAPDH in BMP2 or BMP7 mRNA in MSCs transfected with either LV-BMP2 or LV-BMP2/7 plasmid. Statistical significance determined by one-way ANOVA with Tukey post hoc analysis. (+/- SD, n = 3). C Western blot of MSCs that were untransfected or transfected with LV-BMP2 or LV-BMP2/7. Actin was used as a loading control. D NanoSight analysis demonstrating mean size profile (+/- SD) of extracellular vesicles isolated from LV-BMP2 or LV-BMP2/7 transfected MSCs. E Transmission electron microscopy (TEM) demonstrating size profile of BMP2-EV or BMP2/7-EV. Scale bar 200 nm. F Western blot of small extracellular vesicles collected from MSCs that were untransfected or transfected with LV-BMP2 or LV-BMP2/7. Samples were probed for small extracellular vesicle markers CD63, Flotillin, and CD9. G Western blot of small extracellular vesicles collected from MSCs that were untransfected or transfected with LV-BMP2 or LV-BMP2/7. Samples were probed for BMP2 and BMP7 with actin as a loading control. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/40461558), licensed under a CC-BY license. Not internally tested by R&D Systems.

Applications for Human CD63 Antibody

Application
Recommended Usage

CyTOF-ready

Ready to be labeled using established conjugation methods. No BSA or other carrier proteins that could interfere with conjugation.

Flow Cytometry

0.25 µg/106 cells
Sample: Human PBMCs fixed and permeabilized with FlowX FoxP3/Transcription Factor Fixation & Perm Buffer Kit (Catalog # FC012); PBMC monocytes (surface stain)

Immunohistochemistry

0.3-25 µg/mL
Sample: Immersion fixed paraffin-embedded sections of human lung cancer tissue

Simple Western

20 µg/mL
Sample: Human platelets

Western Blot

0.5 µg/mL
Sample: Human platelets

Reviewed Applications

Read 1 review rated 5 using MAB50482 in the following applications:

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Advanced Features

  • Spectra Viewer - Custom analysis of spectra from multiple fluorochromes
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  • Antigen Density Selector - Match fluorochrome brightness with antigen density
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Flow Cytometry

Formulation, Preparation, and Storage

Purification

Protein A or G purified from cell culture supernatant

Reconstitution

Reconstitute at 0.5 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.

Formulation

Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.

Shipping

Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.

Stability & Storage

Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 12 months from date of receipt, -20 to -70 °C as supplied.
  • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
  • 6 months, -20 to -70 °C under sterile conditions after reconstitution.

Calculators

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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Background: CD63

CD63, also known as LAMP-3 or ME491 (melanoma-associated antigen), is a 30-60 kDa member of the tetraspanin superfamily of protein trafficking proteins. CD63 is ubiquitously expressed and found in late endocytic vesicles, but following cell activation is also present on the plasma membrane. Interaction of CD63 with other membrane proteins or adaptors regulates cell activities such as adhesion, migration and degranulation. Extracellular regions of human CD63 share 67% and 65% amino acid sequence identity with mouse and rat CD63, respectively.

Alternate Names

CD63, Granulophysin, Lamp-3, ME491, OMA81H, Tspan30

Entrez Gene IDs

967 (Human); 12512 (Mouse); 29186 (Rat); 474391 (Canine)

Gene Symbol

CD63

UniProt

P08962-1

Additional CD63 Products

Product Documents for Human CD63 Antibody

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot or batch number in the search box below.

Note: Certificate of Analysis not available for kit components.

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Product Specific Notices for Human CD63 Antibody

For research use only

Citations for Human CD63 Antibody

Customer Reviews for Human CD63 Antibody (1)

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  • Human CD63 Antibody
    Name: Anonymous
    Application: Immunohistochemistry
    Sample Tested: Uterine epithelium
    Species: Human
    Verified Customer | Posted 11/25/2021
    Human CD63 Antibody MAB50482

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