Hi, does this antibody work in western blots under reducing conditions? Thanks!

CyTOF-ready, Flow Cytometry, Immunocytochemistry/ Immunofluorescence, Immunohistochemistry, Immunohistochemistry-Paraffin, Immunoprecipitation, Western Blot

Cited:

Flow Cytometry, IF/IHC, Immunocytochemistry/ Immunofluorescence, Immunohistochemistry, Western Blot

Label

Unconjugated

Antibody Source

Monoclonal Mouse IgG₁ Clone # MEM-259

Format

BSA Free

Concentration

1.0 mg/ml

View all CD63 Antibodies »

Product Specifications

Immunogen

HPB-ALL T cell line

Reactivity Notes

Use in Mouse and Rat reported in scientific literature (PMID:32111836).

Specificity

The antibody MEM-259 reacts with an extracellular/luminal epitope of CD63 (LAMP-3), a 40-60 kDa tetraspan glycoprotein expressed by granulocytes, platelets, T-cells, monocytes/macrophages and endothelial cells. Cell surface exposition of CD63 is usually activation-dependent.

Marker

Late Endosomes Marker

Clonality

Monoclonal

Host

Mouse

Isotype

IgG₁

Theoretical MW

60 kDa.
Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.

Scientific Data Images for CD63 Antibody (MEM-259) - BSA Free

Immunocytochemistry/ Immunofluorescence: CD63 Antibody (MEM-259) - BSA Free [NB100-77913]

Immunocytochemistry/Immunofluorescence: CD63 Antibody (MEM-259) [NB100-77913] - Human HeLa cells. ICC/IF image submitted by a verified customer review.

Western Blot: CD63 Antibody (MEM-259)BSA Free [NB100-77913]

Western Blot: CD63 Antibody (MEM-259) [NB100-77913] - Immunoblot analysis of CD63, CD81, TSG101, and CD9 protein in LSC-Exo and MSC-Exo. Image collected and cropped by CiteAb from the following publication (nature.com/articles/s41467-020-14344-7), licensed under a CC-BY license.

Immunohistochemistry-Paraffin: CD63 Antibody (MEM-259) - BSA Free [NB100-77913]

Immunohistochemistry-Paraffin: CD63 Antibody (MEM-259) [NB100-77913] - Staining of human spleen (paraffin sections) using anti-CD63 (MEM-259).

Flow Cytometry: CD63 Antibody (MEM-259) - BSA Free [NB100-77913]

Flow Cytometry: CD63 Antibody (MEM-259) [NB100-77913] - Flow Cytometry: CD63 Antibody (MEM-259) [FITC] [NB500-483] - Analysis of IgE-activated peripheral blood stained with anti-human CD63 (MEM-259) PE-CyTM5. Image using the FITC form of this antibody.

Immunocytochemistry/ Immunofluorescence: CD63 Antibody (MEM-259) - BSA Free [NB100-77913]

Immunocytochemistry/Immunofluorescence: CD63 Antibody (MEM-259) [NB100-77913] - Staining of human skin fibroblasts with anti-CD63 (MEM-259; green) after co-incubation of living cells with human Transferrin - Dyomics 547 (red); cell nuclei stained with DAPI (blue).

Immunocytochemistry/ Immunofluorescence: CD63 Antibody (MEM-259) - BSA Free [NB100-77913]

Immunocytochemistry/Immunofluorescence: CD63 Antibody (MEM-259) [NB100-77913] - Staining of CD63 in human primary fibroblasts using anti-CD63 (MEM-259; green). Actin cytoskeleton was decorated by phalloidin (red) and cell nuclei stained with DAPI (blue).

Immunocytochemistry/ Immunofluorescence: CD63 Antibody (MEM-259) - BSA Free [NB100-77913]

Immunocytochemistry/Immunofluorescence: CD63 Antibody (MEM-259) [NB100-77913] - Staining of CD63 in human HeLa cell line using anti-CD63 (MEM-259; green). Actin cytoskeleton was decorated by phalloidin (red) and cell nuclei stained with DAPI (blue).

Flow Cytometry: CD63 Antibody (MEM-259) - BSA Free [NB100-77913]

Flow Cytometry: CD63 Antibody (MEM-259) [NB100-77913] - Analysis using the FITC conjugate of NB100-77913. Staining of peripheral blood lymphocytes from a patient with allergy to bee venom after stimulation with bee venom, stained with anti-human CD63 (MEM-259) FITC.

Flow Cytometry: CD63 Antibody (MEM-259) - BSA Free [NB100-77913]

Flow Cytometry: CD63 Antibody (MEM-259) [NB100-77913] - Analysis of IgE-activated peripheral blood stained with anti-human CD63 (MEM-259) PE.

Flow Cytometry: CD63 Antibody (MEM-259) - BSA Free [NB100-77913]

Flow Cytometry: CD63 Antibody (MEM-259) [NB100-77913] - Analysis of IgE-activated peripheral blood stained with anti-human CD63 (MEM-259) purified, GAM-APC.

Flow Cytometry: CD63 Antibody (MEM-259) - BSA Free [NB100-77913]

Flow Cytometry: CD63 Antibody (MEM-259) [NB100-77913] - Analysis of IgE-activated peripheral blood stained with anti-human CD63 (MEM-259) PerCP-CyTM5.5.

Flow Cytometry: CD63 Antibody (MEM-259) - BSA Free [NB100-77913]

Flow Cytometry: CD63 Antibody (MEM-259) [NB100-77913] - Analysis of IgE-activated peripheral blood stained with anti-human CD63 (MEM-259) PerCP.

Flow Cytometry: CD63 Antibody (MEM-259) - BSA Free [NB100-77913]

Flow Cytometry: CD63 Antibody (MEM-259) [NB100-77913] - Flow Cytometry: CD63 Antibody (MEM-259) [FITC] [NB500-483] - Analysis of IgE-activated peripheral blood stained with anti-human CD63 (MEM-259) Alexa Fluor® 488. Image using the FITC form of this antibody.

Western Blot: CD63 Antibody (MEM-259) - BSA Free [NB100-77913] -

Characterization & internalization of M2-Exos. (A) TEM image of M2-Exos. Scale bar: 200 nm. (B) DLS measurement of M2-Exos size. (C) Western blotting assay of exosomal markers in THP-1-M2 cellular lysate & M2-Exos preparation. (D,F) Fluorescence images of 786-O & ACHN cells treated with or without PKH67-labeled M2-Exos (green). Scale bar: 50 μm. (E,G) Three-dimensional confocal reconstruction of 786-O & ACHN cells treated with PKH67-labeled M2-Exos (green). (H) Fluorescence staining analyzing the internalization of M2-Exos by 786-O cells over 12 h. Scale bar: 10 μm. CT: control. Image collected & cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/35328425), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: CD63 Antibody (MEM-259) - BSA Free [NB100-77913] -

Western Blot: CD63 Antibody (MEM-259) - BSA Free [NB100-77913] - Cytokine & checkpoint molecule arrays from plasma exosomes showed a decreased concentration of IFN-gamma, IL-10, IL-13, B7-1, B7-2, & ICOSL in GBM patients in comparison to normal donors. (A) Quantification of IFN-gamma, IL-10, & IL-13 in plasma exosomes from normal donors & grade 4 GBM patients' plasma exosomes (mean ± SEM, n = 4/group, *P < 0.05). (B) Quantification of B7-1, B7-2, & ICOSL in plasma exosomes from normal donors & grade 4 GBM patients' plasma exosomes (mean ± SEM, n = 4/group). (C) Quantification of the immunosuppressive checkpoint & T cell costimulatory homolog protein PD-L1 in plasma exosomes from normal donors & grade 4 GBM patients' plasma exosomes (mean ± SEM, n = 4/group). Interestingly, PD-L1 is found at similar levels in both normal donor & glioma patient plasma exosomes & these findings were confirmed by western blot. Western Blot analysis also shows that exosomal markers Flotillin-1 & CD63 are found universally in plasma exosomes from normal donors (n = 3), glioblastoma (grade 4) patients (n = 4), & grade 3 glioma patients (n = 3), though CD63 expression may be slightly reduced in glioma patients. Image collected & cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/31380286), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

View 12 more images

Applications for CD63 Antibody (MEM-259) - BSA Free

Application

Recommended Usage

Flow Cytometry

2 ug/ml

Immunocytochemistry/ Immunofluorescence

1:10 - 1:2000

Immunohistochemistry

10 ug/mL

Immunohistochemistry-Paraffin

10 ug/ml

Immunoprecipitation

1:10 - 1:500

Western Blot

1:100 - 1:2000

Application Notes

This antibody is CyTOF ready. WB reactivity reported in (PMID: 25034888). Clone MEM-259 has been used for extracellular vesicle flow cytometry

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

Reviewed Applications

Read 1 review rated 5 using NB100-77913 in the following applications:

Immunofluorescence (1 Review)

Formulation, Preparation, and Storage

Purification

Protein A purified

Formulation

PBS (pH 7.4)

Format

BSA Free

Preservative

0.9% Sodium Azide

Concentration

1.0 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. Do not freeze.

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

Entrez Gene IDs

967 (Human)

Gene Symbol

CD63

UniProt

P08962

Additional CD63 Products

Product Documents for CD63 Antibody (MEM-259) - BSA Free

Product Datasheets

Download Product Datasheets

COA

SDS

Download SDS

Product Specific Notices for CD63 Antibody (MEM-259) - 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.

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CD63 Antibody (MEM-259) - BSA Free

Novus Biologicals, part of Bio-Techne | Catalog # NB100-77913

Key Product Details

Species Reactivity

Validated:

Cited:

Applications

Validated:

Cited:

Label

Antibody Source

Format

Concentration

View all CD63 Antibodies »

Product Specifications

Immunogen

Reactivity Notes

Specificity

Marker

Clonality

Host

Isotype

Theoretical MW

Scientific Data Images for CD63 Antibody (MEM-259) - BSA Free

Immunocytochemistry/ Immunofluorescence: CD63 Antibody (MEM-259) - BSA Free [NB100-77913]

Western Blot: CD63 Antibody (MEM-259)BSA Free [NB100-77913]

Immunohistochemistry-Paraffin: CD63 Antibody (MEM-259) - BSA Free [NB100-77913]

Flow Cytometry: CD63 Antibody (MEM-259) - BSA Free [NB100-77913]

Immunocytochemistry/ Immunofluorescence: CD63 Antibody (MEM-259) - BSA Free [NB100-77913]

Immunocytochemistry/ Immunofluorescence: CD63 Antibody (MEM-259) - BSA Free [NB100-77913]

Immunocytochemistry/ Immunofluorescence: CD63 Antibody (MEM-259) - BSA Free [NB100-77913]

Flow Cytometry: CD63 Antibody (MEM-259) - BSA Free [NB100-77913]

Flow Cytometry: CD63 Antibody (MEM-259) - BSA Free [NB100-77913]

Flow Cytometry: CD63 Antibody (MEM-259) - BSA Free [NB100-77913]

Flow Cytometry: CD63 Antibody (MEM-259) - BSA Free [NB100-77913]

Flow Cytometry: CD63 Antibody (MEM-259) - BSA Free [NB100-77913]

Flow Cytometry: CD63 Antibody (MEM-259) - BSA Free [NB100-77913]

Western Blot: CD63 Antibody (MEM-259) - BSA Free [NB100-77913] -

Western Blot: CD63 Antibody (MEM-259) - BSA Free [NB100-77913] -

Applications for CD63 Antibody (MEM-259) - BSA Free

Flow Cytometry

Immunocytochemistry/ Immunofluorescence

Immunohistochemistry

Immunohistochemistry-Paraffin

Immunoprecipitation

Western Blot

Reviewed Applications

Formulation, Preparation, and Storage

Purification

Formulation

Format

Preservative

Concentration

Shipping

Stability & Storage

Background: CD63

Alternate Names

Entrez Gene IDs

Gene Symbol

UniProt

Additional CD63 Products

Product Documents for CD63 Antibody (MEM-259) - BSA Free

Product Datasheets

COA

Certificate of Analysis

SDS

Product Specific Notices for CD63 Antibody (MEM-259) - BSA Free