Exbio — Research products — Antibodies — CD and related antigens — Anti-Hu CD63 PE

Anti-Hu CD63 PE

Regulatory status

RUO

Antigen

CD63

Clone

MEM-259

Format

Reactivity

Human

Application

FC (QC tested)

Excitation laser

blue (488 nm)

Variant

100 tests

1P-343-T100

In stock

242.00 USD

25 tests

1P-343-T025

Delivery 1 week

121.00 USD

Variant

100 tests

1P-343-T100

In stock

242.00 USD

25 tests

1P-343-T025

Delivery 1 week

121.00 USD

Contact distributor

Product details

Description

Images

References

SDS download

Isotype

Mouse IgG1

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.

Application

FC (QC tested)

Application details

Flow cytometry: The reagent is designed for analysis of human blood cells using 20 μl reagent / 100 μl of whole blood or 10⁶ cells in a suspension. The content of a vial (2 ml) is sufficient for 100 tests.

Reactivity

Human

Immunogen

HPB-ALL T cell line

Preparation

Purified antibody is conjugated with R-phycoerythrin (PE) under optimum conditions. Unconjugated antibody and free fluorochrome are removed by size-exclusion chromatography.

Formulation

Stabilizing phosphate buffered saline (PBS), pH 7.4, 15 mM sodium azide

Storage and handling

Store at 2-8°C. Protect from prolonged exposure to light. Do not freeze.

The product is intended For Research Use Only. Diagnostic or therapeutic applications are strictly forbidden. Products shall not be used for resale or transfer to third parties either as a stand-alone product or as a manufacture component of another product without written consent of EXBIO Praha, a.s. EXBIO Praha, a.s. will not be held responsible for patent infringement or any other violations of intellectual property rights that may occur with the use of the products. Orders for all products are accepted subject to the Term and Conditions available at www.exbio.cz. EXBIO, EXBIO Logo, and all other trademarks are property of EXBIO Praha, a.s.

Other names

OMA81H, Granulophysin, Tetraspanin-30, Tspan-30, MLA1, ME491, LAMP-3, OMA81H, TSPAN30

Antigen description

CD63 (LAMP-3, lysosome-associated membrane protein-3), a glycoprotein of tetraspanin family, is present in late endosomes, lysosomes and secretory vesicles of various cell types. It is also present in the plasma membrane, usually following cell activation. Hence, it has become an widely used basophil activation marker. In mast cells, however, CD63 exposition does not need their activation. CD63 interacts with integrins and affects phagocytosis and cell migration, it is also involved in H/K-ATPase trafficking regulation of ROMK1 channels. CD63 also serves as a T-cell costimulation molecule. Expression of CD63 can be used for predicting the prognosis in earlier stages of carcinomas.

Entrez Gene ID 967

UniProt ID P08962

Flow cytometry surface staining pattern of anti-IgE stimulated human peripheral whole blood stained using anti-human CD63 (MEM-259) PE antibody (concentration in sample 1 μg/ml).

Expression profiling on peripheral blood subsets using Anti-human CD63 PE antibody (clone MEM-259). Adaptive panel

HCDM CDMaps standardized procedures (Kuzilkova D et al. Front Immunol. 2022;13:827898) were used for cell isolation and surface staining of blood leukocytes, with the modification of staining protocol using cytometry test tubes.
Suspension of blood leukocytes isolated from buffy coats (2 x 106 cells) was added to the mixture of anti-human CD63 PE antibody (clone MEM-259, 4 µg/ml in stained blood sample) and Monocyte Blocking Buffer (#ED7747), vortexed and incubated for 20 min. Next, optimized backbone antibody panel (HLDA Adaptive) was added to test tubes, vortexed and incubated for 20 min. The residual erythrocytes were lysed with 2 ml of 10× diluted EXCELLYSE Easy solution (#ED7066) and incubated for 10 min. Finally, samples were centrifuged (670 g, 5 min.), supernatant removed and the cell pellet was resuspended in 200 µl of PBS for acquisition.

1P-343_FC_CDMaps-histogram-adaptive-intra

Expression profiling on peripheral blood subsets using Anti-human CD63 PE antibody (clone MEM-259). Intracellular staining, adaptive panel

Analysis of the antibody staining profile was performed on blood leukocytes isolated from buffy coats and permeabilized using EXCELLYSE XPerm buffer set (#ED7397).
HCDM CDMaps standardized procedures (Kuzilkova D et al. Front Immunol. 2022;13:827898) were used for leukocyte isolation.
Suspension of blood leukocytes isolated from buffy coats (2 x 10⁶ cells) was added to the mixture of optimized backbone antibody panel (HLDA Adaptive) and Monocyte Blocking Buffer (#ED7747), vortexed and incubated for 20 min.
Next, EXCELLYSE XPerm buffer set (#ED7397) was used for leukocytes permeabilization according to the instruction procedure.
Mouse monoclonal anti-human CD63 PE antibody (clone MEM-259) was used in concentration 4 µg/ml in stained blood sample (2 x 10⁶ cells).

Expression profiling on peripheral blood subsets using Anti-human CD63 PE antibody (clone MEM-259). Innate panel

HCDM CDMaps standardized procedures (Kuzilkova D et al. Front Immunol. 2022;13:827898) were used for cell isolation and surface staining of blood leukocytes, with the modification of staining protocol using cytometry test tubes.
Suspension of blood leukocytes isolated from buffy coats (2 x 106 cells) was added to the mixture of anti-human CD63 PE antibody (clone MEM-259, 4 µg/ml in stained blood sample) and Monocyte Blocking Buffer (#ED7747), vortexed and incubated for 20 min. Next, optimized backbone antibody panel (HLDA Innate) was added to test tubes, vortexed and incubated for 20 min. The residual erythrocytes were lysed with 2 ml of 10× diluted EXCELLYSE Easy solution (#ED7066) and incubated for 10 min. Finally, samples were centrifuged (670 g, 5 min.), supernatant removed and the cell pellet was resuspended in 200 µl of PBS for acquisition.

Expression profiling on peripheral blood subsets using Anti-human CD63 PE antibody (clone MEM-259). Intracellular staining, innate panel

Analysis of the antibody staining profile was performed on blood leukocytes isolated from buffy coats and permeabilized using EXCELLYSE XPerm buffer set (#ED7397).
HCDM CDMaps standardized procedures (Kuzilkova D et al. Front Immunol. 2022;13:827898) were used for leukocyte isolation.
Suspension of blood leukocytes isolated from buffy coats (2 x 10⁶ cells) was added to the mixture of optimized backbone antibody panel (HLDA Innate) and Monocyte Blocking Buffer (#ED7747), vortexed and incubated for 20 min.
Next, EXCELLYSE XPerm buffer set (#ED7397) was used for leukocytes permeabilization according to the instruction procedure.
Mouse monoclonal anti-human CD63 PE antibody (clone MEM-259) was used in concentration 4 µg/ml in stained blood sample (2 x 10⁶ cells).

Anti-human CD63 PE antibody (clone MEM-259) works in flow cytometry application.

Analysis of the antibody staining profile was performed on blood leukocytes isolated from buffy coats. HCDM CDMaps standardized procedures (Kuzilkova D et al. Front Immunol. 2022;13:827898) were used for cell isolation and surface staining of blood leukocytes, with the modification of staining protocol using cytometry test tubes.
Mouse monoclonal anti-human CD63 PE antibody (clone MEM-259) was used in concentration 4 µg/ml in stained blood sample (2 x 10⁶ cells).

Flow cytometry multicolor surface staining pattern of anti-IgE stimulated human peripheral blood mononuclear cells stained using anti-human CD203c (UCHT1) APC antibody (10 μl reagent / 100 μl of peripheral whole blood) and anti-human CD63 (MEM-259) PE antibody (concentration in sample 1 μg/ml).

Separation of human CD63 positive CD203c positive basophils (red-filled) from CD63 negative lymphocytes (black-dashed) in flow cytometry analysis (surface staining) of anti-IgE stimulated human peripheral whole blood stained using anti-human CD63 (MEM-259) PE antibody (concentration in sample 1 μg/ml).

General references:

Lin D, Kamsteeg EJ, Zhang Y, Jin Y, Sterling H, Yue P, Roos M, Duffield A, Spencer J, Caplan M, Wang WH: Expression of tetraspan protein CD63 activates protein tyrosine kinase (PTK) and enhances the PTK-induced inhibition of ROMK channels. J Biol Chem. 2008 Jan 22
PubMed

Kwon MS, Shin SH, Yim SH, Lee KY, Kang HM, Kim TM, Chung YJ: CD63 as a biomarker for predicting the clinical outcomes in adenocarcinoma of lung. Lung Cancer. 2007 Jul;57(1):46-53.
PubMed

Israels SJ, McMillan-Ward EM: CD63 modulates spreading and tyrosine phosphorylation of platelets on immobilized fibrinogen. Thromb Haemost. 2005 Feb;93(2):311-8.
PubMed

Pfistershammer K, Majdic O, Stöckl J, Zlabinger G, Kirchberger S, Steinberger P, Knapp W: CD63 as an activation-linked T cell costimulatory element. J Immunol. 2004 Nov 15;173(10):6000-8.
PubMed

Mantegazza AR, Barrio MM, Moutel S, Bover L, Weck M, Brossart P, Teillaud JL, Mordoh J: CD63 tetraspanin slows down cell migration and translocates to the endosomal-lysosomal-MIICs route after extracellular stimuli in human immature dendritic cells. Blood. 2004 Aug 15;104(4):1183-90.
PubMed

Grützkau A, Smorodchenko A, Lippert U, Kirchhof L, Artuc M, Henz BM: LAMP-1 and LAMP-2, but not LAMP-3, are reliable markers for activation-induced secretion of human mast cells. Cytometry A. 2004 Sep;61(1):62-8.
PubMed

Product specific references:

Heneberg P, Riegerová K, Kučera P: Pimecrolimus is a potent inhibitor of allergic reactions to hymenopteran venom extracts and birch pollen allergen in vitro. PLoS One. 2015 Nov 12;10(11):e0142953.
PubMed

Cerny J, Feng Y, Yu A, Miyake K, Borgonovo B, Klumperman J, Meldolesi J, McNeil PL, Kirchhausen T: The small chemical vacuolin-1 inhibits Ca(2+)-dependent lysosomal exocytosis but not cell resealing. EMBO Rep. 2004 Sep;5(9):883-8. Erratum in: EMBO Rep. 2005 Sep;6(9):898.
PubMed