Western Blot, ELISA, Flow Cytometry, Flow (Intracellular), Immunocytochemistry/ Immunofluorescence, Immunoprecipitation, Dot Blot, Functional, In vitro assay, Electron Microscopy, CyTOF-ready

Label

Unconjugated

Antibody Source

Monoclonal Mouse IgG₁ kappa Clone # H5C6

Format

Azide and BSA Free

Concentration

1 mg/ml

View all CD63 Antibodies »

Product Specifications

Immunogen

Human splenic adherent cells.

Clonality

Monoclonal

Host

Mouse

Isotype

IgG₁ kappa

Scientific Data Images for CD63 Antibody (H5C6) - Azide and BSA Free

Western Blot: CD63 Antibody (H5C6)Azide and BSA Free [NBP2-80654]

Western Blot: CD63 Antibody (H5C6) - Azide and BSA Free [NBP2-80654] - THP1 whole cell protein was separated by SDS-PAGE on a 12% gel and transferred to PVDF membrane. The membrane was probed with anti-CD63 antibody at 2 ug/ml and detected with an anti-mouse HRP secondary antibody using chemiluminescence. Image from the standard format of this antibody.

Immunocytochemistry/ Immunofluorescence: CD63 Antibody (H5C6) - Azide and BSA Free [NBP2-80654]

Immunocytochemistry/Immunofluorescence: CD63 Antibody (H5C6) - Azide and BSA Free [NBP2-80654] - The CD63 (H5C6) antibody was tested in HeLa cells at a 1:50 dilution against DyLight 488 (Green). Actin and nuclei were counterstained against Phalloidin 568 (Red) and DAPI (Blue), respectively. Image from the standard format of this antibody.

Flow Cytometry: CD63 Antibody (H5C6) - Azide and BSA Free [NBP2-80654]

Flow Cytometry: CD63 Antibody (H5C6) - Azide and BSA Free [NBP2-80654] - Human peripheral blood cells were stained (2 x 10^6 cells/ml) using the anti-CD63 antibody (Blue) at a dilution of 1:1000. Signal was detected using a Gt x Ms DyLight 488 Secondary and gated to the monocyte/granulocyte cell populations. Isotype was Mouse IgG1 kappa (orange). Data collected on BD FACS Calibur flow cytometer. Image from the standard format of this antibody.

Western Blot: CD63 Antibody (H5C6)Azide and BSA Free [NBP2-80654]

Western Blot: CD63 Antibody (H5C6) - Azide and BSA Free [NBP2-80654] - The same samples and volumes were run under reducing and non-reducing conditions. All procedures were performed in parallel for both conditions. L- ladder +- exosomes. Image visualized with HRP linked to secondary antibody. This image was submitted via customer Review. Image from the standard format of this antibody.

Immunocytochemistry/ Immunofluorescence: CD63 Antibody (H5C6) - Azide and BSA Free [NBP2-80654]

Immunocytochemistry/Immunofluorescence: CD63 Antibody (H5C6) - Azide and BSA Free [NBP2-80654] - HEK cells stained with CD63 antibody at a dilution of 1:50 followed by Donkey anti-mouse secondary antibody conjugated with Alexa Fluor 488 (1:500). Nuclei were stained with Hoechst 33342. Image from verified customer review. Image from the standard format of this antibody.

Immunocytochemistry/ Immunofluorescence: CD63 Antibody (H5C6) - Azide and BSA Free [NBP2-80654]

Immunocytochemistry/Immunofluorescence: CD63 Antibody (H5C6) - Azide and BSA Free [NBP2-80654] - HeLa cells were fixed for 10 minutes using 10% formalin and then permeabilized for 5 minutes using 1X PBS + 0.05% Triton X-100. The cells were incubated with anti-CD63 [H5C6] conjugated to Alexa Fluor 488 [NBP2-42225AF488] at 10ug/ml for 1 hour at room temperature. Nuclei were counterstained with DAPI (Blue). Cells were imaged using a 40X objective. Image from the standard format of this antibody.

Immunocytochemistry/ Immunofluorescence: CD63 Antibody (H5C6) - Azide and BSA Free [NBP2-80654]

Immunocytochemistry/Immunofluorescence: CD63 Antibody (H5C6) - Azide and BSA Free [NBP2-80654] - MDCK cells stained with CD63 antibody at a dilution of 1:50 followed by Donkey anti-mouse secondary antibody conjugated with Alexa Fluor 488 (1:500). Nuclei were stained with Hoechst 33342. Image from verified customer review. Image from the standard format of this antibody.

Flow Cytometry: CD63 Antibody (H5C6) - Azide and BSA Free [NBP2-80654]

Flow Cytometry: CD63 Antibody (H5C6) - Azide and BSA Free [NBP2-80654] - A431 cells were stained (1 x10^6 cells/ml) using the anti-CD63 antibody at a 1:1000 dilution (blue). Signal was detected with Gt x Ms Dylight 488 secondary. Isotype control (orange). Image from the standard format of this antibody.

Flow Cytometry: CD63 Antibody (H5C6) - Azide and BSA Free [NBP2-80654]

Flow Cytometry: CD63 Antibody (H5C6) - Azide and BSA Free [NBP2-80654] - An intracellular stain was performed on HeLa cells with CD63 [H5C6] Antibody NBP2-42225AF488 (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 directly conjugated to Alexa Fluor 488.

Flow Cytometry: CD63 Antibody (H5C6) - Azide and BSA Free [NBP2-80654]

Flow Cytometry: CD63 Antibody (H5C6) - Azide and BSA Free [NBP2-80654] - An intracellular stain was performed on HeLa cells with CD63 Antibody (H5C6) NBP2-42225APC (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 1 ug/mL for 30 minutes at room temperature. Both antibodies were directly conjugated to Allophycocyanin.

Flow Cytometry: CD63 Antibody (H5C6) - Azide and BSA Free [NBP2-80654]

Flow Cytometry: CD63 Antibody (H5C6) - Azide and BSA Free [NBP2-80654] - An intracellular stain was performed on SK-MEL-28 cells with CD63 Antibody (H5C6) NBP2-42225AF488 (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 directly conjugated to Alexa Fluor 488.

View 8 more images

Applications for CD63 Antibody (H5C6) - Azide and BSA Free

Application

Recommended Usage

Electron Microscopy

reported in scientific literature (PMID 16735575)

Flow Cytometry

1:1000

Functional

reported in scientific literature (PMID 9811687)

Immunocytochemistry/ Immunofluorescence

1:50-1:100

In vitro assay

reported in scientific literature (PMID 21464080)

Formulation, Preparation, and Storage

Purification

Protein G purified

Formulation

PBS

Format

Azide and BSA Free

Preservative

No Preservative

Concentration

1 mg/ml

Shipping

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

Stability & Storage

Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.

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) - Azide and BSA Free

Product Datasheets

Download Product Datasheets

COA

SDS

Download SDS

Product Specific Notices for CD63 Antibody (H5C6) - Azide and BSA Free

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.

Citations
Reviews

CD63 Antibody (H5C6) - Azide and BSA Free

Novus Biologicals, part of Bio-Techne | Catalog # NBP2-80654

Key Product Details

Species Reactivity

Applications

Label

Antibody Source

Format

Concentration

View all CD63 Antibodies »

Product Specifications

Immunogen

Clonality

Host

Isotype

Scientific Data Images for CD63 Antibody (H5C6) - Azide and BSA Free

Western Blot: CD63 Antibody (H5C6)Azide and BSA Free [NBP2-80654]

Immunocytochemistry/ Immunofluorescence: CD63 Antibody (H5C6) - Azide and BSA Free [NBP2-80654]

Flow Cytometry: CD63 Antibody (H5C6) - Azide and BSA Free [NBP2-80654]

Western Blot: CD63 Antibody (H5C6)Azide and BSA Free [NBP2-80654]

Immunocytochemistry/ Immunofluorescence: CD63 Antibody (H5C6) - Azide and BSA Free [NBP2-80654]

Immunocytochemistry/ Immunofluorescence: CD63 Antibody (H5C6) - Azide and BSA Free [NBP2-80654]

Immunocytochemistry/ Immunofluorescence: CD63 Antibody (H5C6) - Azide and BSA Free [NBP2-80654]

Flow Cytometry: CD63 Antibody (H5C6) - Azide and BSA Free [NBP2-80654]

Flow Cytometry: CD63 Antibody (H5C6) - Azide and BSA Free [NBP2-80654]

Flow Cytometry: CD63 Antibody (H5C6) - Azide and BSA Free [NBP2-80654]

Flow Cytometry: CD63 Antibody (H5C6) - Azide and BSA Free [NBP2-80654]

Applications for CD63 Antibody (H5C6) - Azide and BSA Free

Electron Microscopy

Flow Cytometry

Functional

Immunocytochemistry/ Immunofluorescence

In vitro assay

Formulation, Preparation, and Storage

Purification

Formulation

Format

Preservative

Concentration

Shipping

Stability & Storage

Background: CD63

Alternate Names

Gene Symbol

Additional CD63 Products

Product Documents for CD63 Antibody (H5C6) - Azide and BSA Free

Product Datasheets

COA

Certificate of Analysis

SDS

Product Specific Notices for CD63 Antibody (H5C6) - Azide and BSA Free