Invitrogen™ CD63 Monoclonal Antibody (H5C6), eFluor™ 660, eBioscience™

Catalog No.	Quantity
50-112-4280	100 Tests
50-063-941	25 Tests

Catalog No. 50-112-4280 Supplier Invitrogen™ Supplier No. 50063942

Description

Mouse Monoclonal Antibody

Description: This H5C6 monoclonal antibody reacts with human CD63, a type III member of the tetraspanin family of transmembrane proteins. CD63 is expressed intracellularly on lysosomes, endosomes, and granules of resting platelets and basophils. However, cell surface expression of CD63 can be detected on activated basophils and platelets, monocytes, macrophages, and granulocytes. This receptor is also expressed on endothelial cells, fibroblasts, and smooth muscle cells. Studies have demonstrated that CD63 associates with integrins (VLA-3 and VLA-6) and TIMP-1 to mediate the allergic response. Applications Reported: This H5C6 antibody has been reported for use in flow cytometric analysis. Applications Tested: This H5C6 antibody has been pre-titrated and tested by flow cytometric analysis of normal human peripheral blood cells. This can be used at 5 μL (0.5 μg) per test. A test is defined as the amount (μg) of antibody that will stain a cell sample in a final volume of 100 μL. Cell number should be determined empirically but can range from 10^5 to 10^8 cells/test. eFluor® 660 is a replacement for Alexa Fluor® 647. eFluor® 660 emits at 659 nm and is excited with the red laser (633 nm). Please make sure that your instrument is capable of detecting this fluorochome. Excitation: 633-647 nm; Emission: 668 nm; Laser: Red Laser. Filtration: 0.2 μm post-manufacturing filtered.

CD63, also known as LAMP-3, is a glycoprotein belonging to the tetraspanin family, primarily located in late endosomes, lysosomes, and secretory vesicles across various cell types. It is expressed intracellularly in resting platelets and basophils, but upon activation, CD63 translocates to the plasma membrane, making it a widely used marker for basophil activation. Unlike basophils, mast cells can expose CD63 without activation. CD63 is expressed on activated platelets, monocytes, macrophages, and granulocytes, as well as endothelial cells, fibroblasts, and smooth muscle cells. It interacts with integrins (such as VLA-3 and VLA-6) and TIMP-1, playing roles in mediating allergic responses, phagocytosis, and cell migration. Additionally, CD63 is involved in the regulation of H/K-ATPase trafficking and ROMK1 channels, and serves as a T-cell costimulation molecule. CD63 expression can be used to predict prognosis in early-stage carcinomas and is identical to the melanoma-associated antigen ME491 and the platelet antigen PTLGP40. Diseases associated with CD63 dysfunction include melanoma and Hermansky-Pudlak Syndrome, highlighting its significance in both immune responses and disease processes.

Specifications

• Antigen: CD63
• Applications: Flow Cytometry
• Classification: Monoclonal
• Clone: H5C6
• Concentration: 5 μL/Test
• Conjugate: eFluor 660
• Formulation: PBS with BSA and 0.09% sodium azide; pH 7.2
• Gene: CD63
• Gene Accession No.: P08962
• Gene Alias: C75951; CD 63; Cd63; CD63 antigen; CD63 antigen (melanoma 1 antigen); Cd63 molecule; CD63 protein; granulophysin; LAMP-3; Lysosomal-associated membrane protein 3; lysosome-associated membrane glycoprotein 3; mast cell antigen AD1; ME491; melanoma 1 antigen; melanoma-associated antigen ME491; melanoma-associated antigen MLA1; MLA1; Ocular melanoma-associated antigen; OMA81H; Tetraspanin-30; TSPAN30; tspan-30
• Gene Symbols: CD63
• Host Species: Mouse
• Purification Method: Affinity chromatography
• Quantity: 100 Tests
• Regulatory Status: RUO
• Primary or Secondary: Primary
• Gene ID (Entrez): 967
• Target Species: Human
• Content And Storage: 4°C, store in dark, DO NOT FREEZE!
• Product Type: Antibody
• Form: Liquid
• Isotype: IgG1 κ

Frequently Asked Questions (FAQs)

I am unable to analyze my cells stained with eFluor Organic Dyes today. What options do I have?

Our options will depend on the samples you are analyzing.

If cell viability is not critical, you can store your stained samples at 4 degrees C or on ice overnight in the dark and analyze the following day.

For samples stained with eFluor organic fluorochromes, we recommend that cells be suspended in 100 uL of Flow Cytometry Staining Buffer (Cat. No. 00-4222) and 100 uL of eBioscience IC Fixation Buffer (Cat. No. 00-8222); samples can be incubated for up to 3 days at 4 degrees C in the dark. Alternatively, the 1-step Fix/Lyse Solution (Cat. No. 00-5333) can be used. This is a great option when working with whole blood but also works for other cell types.

Can the eFluor Organic fluorochromes be used for intracellular staining?

Yes, the eFluor Organic fluorochromes can be used for intracellular staining. The eFluor organic fluorochromes maintain bright signal and require minimal changes in compensation when fixed with eBioscience IC Fixation Buffer (Cat. No. 00-8222-49) and Permeabilization Buffer (Cat. No. 00-8333-56) or 1-step Fix/Lyse Solution (Cat. No. 00-5333-54, 00-5333-57) (as compared to live cells).

Is the eFluor 660 fluorochrome compatible with Anti-Cy5/Alexa Fluor 647 beads?

Yes, in-house studies have demonstrated that the eFluor 660 fluorochrome is recognized by Anti-Cy5/Alexa Fluor 647 beads. Side by side studies with Alexa Fluor 647 versus eFluor 660 conjugated antibodies have demonstrated comparable results.

What buffers are compatible with the eFluor Organic fluorochromes and eVolve QDots?

The eFluor Organic fluorochromes and eVolve QDots can be used with flow staining buffers containing PBS and protein.

What is the difference between eFluor Organic Dyes and eVolve Qdots?

The eFluor Organic Dyes (eFluor 450, APC-eFluor 780, PerCP-eFluor 710, eFluor 710) are conventional fluorochromes. In contrast, the eVolve line of products are Quantum dots.

Are the eFluor Organic Dyes photo-labile?

As with other fluorochromes, we recommend minimal exposure to light to maintain optimal signal.