Anti-CD63 antibody [EPR5702] - Late Endosome Marker is a rabbit recombinant monoclonal antibody that is used to detect CD63 in IHC-P, Western blot. Suitable for Human samples.
- Using biophysical QC, antibody identity is confirmed at a molecular level for unrivaled batch-batch consistency
- Specificity confirmed with CD63 knockout cell line validation
- Specificity and sensitivity confirmed in IHC with multi-tissue microarray (TMA) validation
- Antibody clone EPR5702 is cited in >540 publications
pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
IHC-P | WB | |
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Human | Tested | Tested |
Species | Dilution info | Notes |
---|---|---|
Species Human | Dilution info 1/2000 | Notes For unpurified use at 1/250 - 1/500. Perform heat-mediated antigen retrieval before commencing with IHC staining protocol. |
Species | Dilution info | Notes |
---|---|---|
Species Human | Dilution info 1/1000 - 1/10000 | Notes - |
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Functions as a cell surface receptor for TIMP1 and plays a role in the activation of cellular signaling cascades. Plays a role in the activation of ITGB1 and integrin signaling, leading to the activation of AKT, FAK/PTK2 and MAP kinases. Promotes cell survival, reorganization of the actin cytoskeleton, cell adhesion, spreading and migration, via its role in the activation of AKT and FAK/PTK2. Plays a role in VEGFA signaling via its role in regulating the internalization of KDR/VEGFR2. Plays a role in intracellular vesicular transport processes, and is required for normal trafficking of the PMEL luminal domain that is essential for the development and maturation of melanocytes. Plays a role in the adhesion of leukocytes onto endothelial cells via its role in the regulation of SELP trafficking. May play a role in mast cell degranulation in response to Ms4a2/FceRI stimulation, but not in mast cell degranulation in response to other stimuli.
CD63, MLA1, TSPAN30, CD63 antigen, Granulophysin, Lysosomal-associated membrane protein 3, Lysosome integral membrane protein 1, Melanoma-associated antigen ME491, OMA81H, Ocular melanoma-associated antigen, Tetraspanin-30, LAMP-3, Limp1, Tspan-30
Anti-CD63 antibody [EPR5702] - Late Endosome Marker is a rabbit recombinant monoclonal antibody that is used to detect CD63 in IHC-P, Western blot. Suitable for Human samples.
- Using biophysical QC, antibody identity is confirmed at a molecular level for unrivaled batch-batch consistency
- Specificity confirmed with CD63 knockout cell line validation
- Specificity and sensitivity confirmed in IHC with multi-tissue microarray (TMA) validation
- Antibody clone EPR5702 is cited in >540 publications
pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Patented technology
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
What are the advantages of a recombinant monoclonal antibody?
This product is a recombinant monoclonal antibody, which offers several advantages including:
For more information, read more on recombinant antibodies.
Species reactivity
Rat: We have preliminary internal testing data to indicate this antibody may not react with this species.
Please contact us for more information.
CD63 is a protein commonly known as LAMP-3 a member of the tetraspanin family with a molecular weight of approximately 25-65 kDa depending on post-translational modifications such as glycosylation. It resides mainly on the membrane of intracellular vesicles such as lysosomes and platelets. CD63 also appears on the surface of activated cells particularly in immune cells. Researchers often use CD63 as a marker in assays including CD63 western blot techniques due to its distinct expression patterns.
CD63 participates in various cellular processes including membrane trafficking cell adhesion and signal transduction. It associates with integrins and other tetraspanins forming complexes that influence cellular adhesion and migration. Through its role in exosome formation CD63 contributes significantly to intercellular communication by facilitating the transfer of proteins lipids and RNA between cells.
CD63 plays a role in pathways linked to cell proliferation and migration and immune response regulation. Within the integrin-mediated signaling pathway CD63 interacts with proteins such as integrins to promote cell adhesion and migration. CD63 also links to vesicle trafficking pathways collaborating with proteins involved in exocytosis and endocytosis which affect cell membrane restructuring and intracellular transport.
CD63 shows connections with cancer and immune system disorders. Its role in promoting cell adhesion and migration links it with cancer metastasis including melanoma and breast cancer where CD63 expression levels often rise. Additionally CD63 associates with allergic reactions due to its involvement in histamine release where it impacts immune proteins like FcεRI which participate in allergy pathogenesis.
We have tested this species and application combination and it works. It is covered by our product promise.
We have not tested this specific species and application combination in-house, but expect it will work. It is covered by our product promise.
This species and application combination has not been tested, but we predict it will work based on strong homology. However, this combination is not covered by our product promise.
We do not recommend this combination. It is not covered by our product promise.
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In the unlikely event of one of our products not working as expected, you are covered by our product promise.
Full details and terms and conditions can be found here:
Terms & Conditions.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human melanoma tissue labelling CD63 with unpurified ab134045 at a dilution of 1/250.
Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
ab134045 was shown to specifically react with CD63 in wild-type HAP1 Brefeldin A treated cells as signal was lost in HAP1 Brefeldin A treated CD63 knockout cells. Wild-type and CD63 knockout samples were subjected to SDS-PAGE. ab134045 and Anti-GAPDH antibody [mAbcam 9484] - Loading Control ab9484 (Mouse anti-GAPDH loading control) were incubated overnight at 4°C at 1/1000 dilution and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.
All lanes: Western blot - Anti-CD63 antibody [EPR5702] - Late Endosome Marker (ab134045) at 1/1000 dilution
Lane 1: Wild-type HAP1 whole cell lysate at 20 µg
Lane 2: Brefeldin A treated wild-type HAP1 whole cell lysate at 20 µg
Lane 3: CD63 knockout HAP1 whole cell lysate at 20 µg
Lane 4: Brefeldin A treated CD63 knockout HAP1 whole cell lysate at 20 µg
Lane 5: HL60 whole cell lysate at 20 µg
Lane 6: Human platelets whole cell lysate at 20 µg
Predicted band size: 26 kDa
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human tonsil tissue labelling CD63 with purified ab134045 at 1/2000. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. Goat Anti-Rabbit IgG H&L (HRP) ab97051, a HRP-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/500). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.
Blocking and dilution buffer: 5% NFDM/TBST.
All lanes: Western blot - Anti-CD63 antibody [EPR5702] - Late Endosome Marker (ab134045) at 1/5000 dilution
Lane 1: A375 whole cell lysate at 20 µg
Lane 2: HL-60 whole cell lysate at 20 µg
Lane 3: Human melanoma tissue lysate at 20 µg
All lanes: HRP-conjugated anti-rabbit IgG, specific to the non-reduced form of IgG at 1/1000 dilution
Predicted band size: 26 kDa
Observed band size: 30-65 kDa
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human skeletal muscle tissue (negative control) labelling CD63 with purified ab134045 at 1/2000. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. Goat Anti-Rabbit IgG H&L (HRP) ab97051, a HRP-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/500). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.
All lanes: Western blot - Anti-CD63 antibody [EPR5702] - Late Endosome Marker (ab134045) at 1/1000 dilution
Lane 1: Human platelet lysate at 10 µg
Lane 2: Human melanoma lysate at 10 µg
Lane 3: HL-60 lysate at 10 µg
Lane 4: A375 lysate at 10 µg
All lanes: HRP-conjugated goat anti-rabbit IgG at 1/2000 dilution
Predicted band size: 26 kDa
Immunohistochemical analysis of formalin fixed paraffin embedded human tonsil staining CD63 with ab134045 at a concentration of 0.1 µg/ml. The immunostaining was performed on a Ventana DISCOVERY ULTRA (Roche Tissue Diagnostics) instrument with a OptiView DAB IHC Detection Kit. Heat mediated antigen retrieval was performed with DISCOVERY cell conditioning solution (CC1) 100°C, pH8.5 for 32mins.
ab134045 anti-CD63 [EPR5702] was incubated for 16mins at 37°C. Sections were counterstained with Hematoxylin II. Image inset shows absence of staining in secondary antibody only control.
Customers are encouraged to optimise antigen retrieval conditions, antibody concentration, incubation times and temperature for best results in their own IHC assay workflow (automated and manual)
Anti-CD63 antibody [EPR5702] (ab134045) staining at 1/1000 dilution, shown in green; Mouse anti-Alpha Tubulin [DM1A] (Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab134045 was shown to bind specifically to CD63. A band was observed at 30 kDa in wild-type HepG2 cell lysates with no signal observed at this size in CD63 knockout cell line Human CD63 knockout Hep G2 cell line ab280796 (knockout cell lysate ab283828). To generate this image, wild-type and CD63 knockout HepG2 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.
All lanes: Western blot - Anti-CD63 antibody [EPR5702] - Late Endosome Marker (ab134045) at 1/1000 dilution
Lanes 1 - 4: Western blot at 20 µg
Lanes 3 - 4: Western blot - Human CD63 knockout Hep G2 cell lysate (ab283828)
Lanes 3 - 4: Western blot - Human CD63 knockout Hep G2 cell line (Human CD63 knockout Hep G2 cell line ab280796)
All lanes: Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution
Performed under reducing conditions.
Predicted band size: 26 kDa
Observed band size: 30 kDa
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