CD63 Antibody (H5C6) [DyLight 550]

Novus Biologicals, part of Bio-Techne | Catalog # NBP2-42225R

Clone H5C6 was used by HLDA to establish CD designation.
Novus Biologicals, part of Bio-Techne
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NBP2-42225R
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Key Product Details

Species Reactivity

Validated:

Human, Canine

Applications

Block/Neutralize, Dot Blot, Electron Microscopy, ELISA, Flow (Intracellular), Flow Cytometry, Functional, Immunocytochemistry/ Immunofluorescence, Immunohistochemistry Whole-Mount, Immunoprecipitation, In vitro assay, Western Blot

Label

DyLight 550 (Excitation = 562 nm, Emission = 576 nm)

Antibody Source

Monoclonal Mouse IgG1 kappa Clone # H5C6

Concentration

Please see the vial label for concentration. If unlisted please contact technical services.

View all CD63 Antibodies »

Product Summary for CD63 Antibody (H5C6) [DyLight 550]

Immunogen

Human splenic adherent cells.

Marker

Exosome Marker

Clonality

Monoclonal

Host

Mouse

Isotype

IgG1 kappa

Scientific Data Images for CD63 Antibody (H5C6) [DyLight 550]

Immunocytochemistry/Immunofluorescence: CD63 Antibody (H5C6) [DyLight 550] [NBP2-42225R] - HeLa cells were fixed in 4% paraformaldehyde for 10 minutes and permeabilized in 0.05% Triton X-100 in PBS for 5 minutes. The cells were incubated with anti-CD63 Antibody [H5C6] conjugated to DyLight 550 (NBP2-42225R) at 5 ug/ml for 1 hour at room temperature. Nuclei were counterstained with DAPI (Blue). Cells were imaged using a 100X objective and digitally deconvolved.
Flow Cytometry: CD63 Antibody (H5C6) [DyLight 550] [NBP2-42225R] - An intracellular stain was performed on SK-MEL-28 cells with CD63 (H5C6) Antibody NBP2-42225R (blue) and a matched isotype control (orange). Cells were fixed with 4% PFA and then permeabilized with 0.1% saponin. Cells were incubated in an antibody dilution of 5 ug/mL for 30 minutes at room temperature. Both antibodies were conjugated to DyLight 550.
CD63 Antibody (H5C6) [DyLight 550]

CD63 (H5C6) in A431 Human Cell Line -

CD63 (H5C6) was detected in immersion fixed A431 human skin carcinoma cell line using Mouse anti-CD63 (H5C6) Protein-G purified Monoclonal Antibody conjugated to DyLight 550 (Catalog # NBP2-42225R) (red) at 5 µg/mL overnight at 4C. Cells were counterstained with DAPI (blue). Cells were imaged using a 100X objective and digitally deconvolved.
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Applications for CD63 Antibody (H5C6) [DyLight 550]

Application
Recommended Usage

Block/Neutralize

Optimal dilutions of this antibody should be experimentally determined.

Dot Blot

Optimal dilutions of this antibody should be experimentally determined.

ELISA

Optimal dilutions of this antibody should be experimentally determined.

Electron Microscopy

Optimal dilutions of this antibody should be experimentally determined.

Flow (Intracellular)

Optimal dilutions of this antibody should be experimentally determined.

Flow Cytometry

Optimal dilutions of this antibody should be experimentally determined.

Functional

Optimal dilutions of this antibody should be experimentally determined.

Immunocytochemistry/ Immunofluorescence

Optimal dilutions of this antibody should be experimentally determined.

Immunohistochemistry Whole-Mount

Optimal dilutions of this antibody should be experimentally determined.

Immunoprecipitation

Optimal dilutions of this antibody should be experimentally determined.

In vitro assay

Optimal dilutions of this antibody should be experimentally determined.

Western Blot

Optimal dilutions of this antibody should be experimentally determined.
Application Notes
Optimal dilution of this antibody should be experimentally determined.
Please Note: Optimal dilutions of this antibody should be experimentally determined.

Formulation, Preparation, and Storage

Purification

Protein G purified

Formulation

50mM Sodium Borate

Preservative

0.05% Sodium Azide

Concentration

Please see the vial label for concentration. If unlisted please contact technical services.

Shipping

The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.

Stability & Storage

Store at 4C in the dark.

Background: CD63

CD63 (cluster of differentiation 63), also known as lysosome associated membrane protein 3 (LAMP-3), is a membrane spanning glycoprotein with a molecular mass ranging from 30-60 kDa that was the first characterized member of the tetraspanin superfamily (1,2). CD63 is ubiquitously expressed and largely present on the cell surface in the endosomal system (1-3). More specifically, it is generally present in multivesicular bodies (MVBs), also called late endosomes, and lysosomes (1). The CD63 molecule is a total of 238 amino acids (aa), has four hydrophobic membrane-spanning regions and three N-glycosylation sites, and the encoded protein has a theoretical molecular weight of 25 kDa (2,3). CD63 both directly and indirectly interacts with other proteins including integrins, cell surface receptors, other tetraspanins, kinases, and adapter proteins, to name a few (1). CD63 is involved in many cell processes including cell survival and activation, cell adhesion, invasion, and migration (1). Additionally, CD63 has been shown to interact with tissue inhibitor of metalloproteinase 1 (TIMP-1), which originally thought to be an inhibitor of cancer progression has recently been shown to have cancer promoting properties as well (1,4). The CD63-TIMP-1 interaction has been shown to activate the PI3K/AKT pathway in lung adenocarcinoma cells and also promote survival and invasion of acute myeloid leukemia cells (1).CD63 is a useful flow cytometry marker for basophil granulocytes and can be used for the basophil activation test (BAT) to assess IgE-mediated allergy response (5). As CD63 was initially discovered on activated blood platelets and is a lysosomal-associated protein it makes sense it would be involved in platelet or lysosomal-related disorders (1,2). More precisely, CD63 is associated with Hermansky-Pudlak syndrome, a disease characterized by albinism and platelet storage pool deficiency (6).

References

1. Pols, M. S., & Klumperman, J. (2009). Trafficking and function of the tetraspanin CD63. Experimental cell research. https://doi.org/10.1016/j.yexcr.2008.09.020

2. Metzelaar, M. J., Wijngaard, P. L., Peters, P. J., Sixma, J. J., Nieuwenhuis, H. K., & Clevers, H. C. (1991). CD63 antigen. A novel lysosomal membrane glycoprotein, cloned by a screening procedure for intracellular antigens in eukaryotic cells. The Journal of biological chemistry.

3. Horejsi, V., & Vlcek, C. (1991). Novel structurally distinct family of leucocyte surface glycoproteins including CD9, CD37, CD53 and CD63. FEBS letters. https://doi.org/10.1016/0014-5793(91)80988-f

4. Eckfeld, C., HauBler, D., Schoeps, B., Hermann, C. D., & Kruger, A. (2019). Functional disparities within the TIMP family in cancer: hints from molecular divergence. Cancer metastasis reviews. https://doi.org/10.1007/s10555-019-09812-6

5. Hoffmann, H. J., Santos, A. F., Mayorga, C., Nopp, A., Eberlein, B., Ferrer, M., Rouzaire, P., Ebo, D. G., Sabato, V., Sanz, M. L., Pecaric-Petkovic, T., Patil, S. U., Hausmann, O. V., Shreffler, W. G., Korosec, P., & Knol, E. F. (2015). The clinical utility of basophil activation testing in diagnosis and monitoring of allergic disease. Allergy. https://doi.org/10.1111/all.12698

6. Dell'Angelica, E. C., Shotelersuk, V., Aguilar, R. C., Gahl, W. A., & Bonifacino, J. S. (1999). Altered trafficking of lysosomal proteins in Hermansky-Pudlak syndrome due to mutations in the beta 3A subunit of the AP-3 adaptor. Molecular cell. https://doi.org/10.1016/s1097-2765(00)80170-7

Alternate Names

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

Gene Symbol

CD63

Additional CD63 Products

Product Documents for CD63 Antibody (H5C6) [DyLight 550]

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Certificate of Analysis

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Product Specific Notices for CD63 Antibody (H5C6) [DyLight 550]



DyLight (R) is a trademark of Thermo Fisher Scientific Inc. and its subsidiaries.

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

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