CD63 Antibody - #AF5117

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Product: CD63 Antibody
Catalog: AF5117
Description: Rabbit polyclonal antibody to CD63
Application: WB IHC
Reactivity: Human, Mouse, Rat
Mol.Wt.: 25kDa,47kDa; 26kD(Calculated).
Uniprot: P08962
RRID: AB_2837603

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Related Downloads
MS Report
HPLC Report
MSDS
Data Sheet
COA
Protocols
WB Handbook
IHC Protocol
IF/ICC Protocol

Product Info

Source:
Rabbit
Application:
IHC 1:50-1:200, WB 1:1000
*The optimal dilutions should be determined by the end user.
*Tips:

WB: For western blot detection of denatured protein samples. IHC: For immunohistochemical detection of paraffin sections (IHC-p) or frozen sections (IHC-f) of tissue samples. IF/ICC: For immunofluorescence detection of cell samples. ELISA(peptide): For ELISA detection of antigenic peptide.

Reactivity:
Human,Mouse,Rat
Clonality:
Polyclonal
Specificity:
CD63 Antibody detects endogenous levels of total CD63.
RRID:
AB_2837603
Cite Format: Affinity Biosciences Cat# AF5117, RRID:AB_2837603.
Conjugate:
Unconjugated.
Purification:
The antiserum was purified by peptide affinity chromatography using SulfoLink™ Coupling Resin (Thermo Fisher Scientific).
Storage:
Rabbit IgG in phosphate buffered saline , pH 7.4, 150mM NaCl, 0.02% sodium azide and 50% glycerol. Store at -20 °C. Stable for 12 months from date of receipt.
Alias:

Fold/Unfold

Lysosomal associated membrane protein 3; CD 63; CD63; CD63 antigen (melanoma 1 antigen); CD63 antigen; CD63 antigen melanoma 1 antigen; CD63 molecule; CD63_HUMAN; gp55; Granulophysin; LAMP 3; LAMP-3; LAMP3; LIMP; 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; Melanoma-associated antigen ME491; MGC72893; MLA 1; MLA1; NGA; Ocular melanoma associated antigen; Ocular melanoma-associated antigen; OMA81H; PTLGP40; Tetraspanin 30; Tetraspanin-30; Tspan 30; Tspan-30; TSPAN30;

Immunogens

Immunogen:
Uniprot:

P08962(CD63_HUMAN) >>Visit HPA database.

Gene(ID):
CD63(967)
Expression:
P08962 CD63_HUMAN:

Detected in platelets (at protein level). Dysplastic nevi, radial growth phase primary melanomas, hematopoietic cells, tissue macrophages.

Description:
Functions as 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/VEGF
Sequence:
MAVEGGMKCVKFLLYVLLLAFCACAVGLIAVGVGAQLVLSQTIIQGATPGSLLPVVIIAVGVFLFLVAFVGCCGACKENYCLMITFAIFLSLIMLVEVAAAIAGYVFRDKVMSEFNNNFRQQMENYPKNNHTASILDRMQADFKCCGAANYTDWEKIPSMSKNRVPDSCCINVTVGCGINFNEKAIHKEGCVEKIGGWLRKNVLVVAAAALGIAFVEVLGIVFACCLVKSIRSGYEVM

PTMs - P08962 As Substrate

Site PTM Type Enzyme
K110 Ubiquitination
K128 Ubiquitination
N130 N-Glycosylation
S159 Phosphorylation
K162 Ubiquitination
K188 Ubiquitination
K194 Ubiquitination

Research Backgrounds

Function:

Functions as 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.

PTMs:

Palmitoylated at a low, basal level in unstimulated platelets. The level of palmitoylation increases when platelets are activated by thrombin (in vitro).

Subcellular Location:

Cell membrane>Multi-pass membrane protein. Lysosome membrane>Multi-pass membrane protein. Late endosome membrane>Multi-pass membrane protein. Endosome>Multivesicular body. Melanosome. Secreted>Extracellular exosome. Cell surface.
Note: Also found in Weibel-Palade bodies of endothelial cells (PubMed:10793155). Located in platelet dense granules (PubMed:7682577). Detected in a subset of pre-melanosomes. Detected on intralumenal vesicles (ILVs) within multivesicular bodies (PubMed:21962903).

Extracellular region or secreted Cytosol Plasma membrane Cytoskeleton Lysosome Endosome Peroxisome ER Golgi apparatus Nucleus Mitochondrion Manual annotation Automatic computational assertionSubcellular location
Tissue Specificity:

Detected in platelets (at protein level). Dysplastic nevi, radial growth phase primary melanomas, hematopoietic cells, tissue macrophages.

Subunit Structure:

Interacts with TIMP1 and ITGB1 and recruits TIMP1 to ITGB1. Interacts with CD9. Identified in a complex with CD9 and ITGB3. Interacts with PMEL. Interacts with KDR/VEGFR2; identified in a complex with ITGB1 and KDR/VEGFR2 and is required to recruit KDR to ITGB1 complexes. Interacts with SYT7 (By similarity).

Family&Domains:

Belongs to the tetraspanin (TM4SF) family.

Research Fields

· Cellular Processes > Transport and catabolism > Lysosome.   (View pathway)

· Human Diseases > Cancers: Overview > Proteoglycans in cancer.

References

1). Exosomes-loaded electroconductive nerve dressing for nerve regeneration and pain relief against diabetic peripheral nerve injury. Bioactive Materials (PubMed: 36923267) [IF=18.9]
2). Iron Oxide Nanoparticles Engineered Macrophage-Derived Exosomes for Targeted Pathological Angiogenesis Therapy. ACS nano (PubMed: 38412252) [IF=17.1]

Application: WB    Species: Mouse    Sample:

Figure 2 Characterization of ESIONPs@EXO derived from ESIONPs engineered macrophages. (A) The morphology of EXO and ESIONPs@EXO determined by TEM. Scale bar: 200 nm (left) and 100 nm (right). (B) The size distribution of EXO and ESIONPs@EXO evaluated by NTA. (C) Western blot analysis of CD9, CD63, CD81, TSG101, and calnexin. (D) Relaxation properties of ESIONPs@EXO. (E) T1 and T2 weighted MR images of ESIONPs@EXO at different concentrations (measured on a 3 T MR scanner). 1/T1 (F) and 1/T2 (G) relaxation rates of ESIONPs@EXO at different concentrations.
3). Exosomal miR-21 from tubular cells contributes to renal fibrosis by activating fibroblasts via targeting PTEN in obstructed kidneys. Theranostics (PubMed: 34522205) [IF=12.4]

Application: WB    Species: Mice    Sample: NRK-52E cells

Figure 2 TGF-β1 promotes the secretion of exosomes by renal tubular epithelial cells and activates fibroblasts in vitro. (A) Schematic diagram of experimental process. Exosomes from NRK-52E cells treated without (Ctrl-Exos) or with TGF-β1 (TGFβ1-Exos) were extracted and incubated with NRK-49F cells. (B,C) DLS and NTA of exosomes from NRK-52E cells. (D) TEM image of exosomes isolated from NRK-52E cells. Scale bar = 100 nm. (E, F) Representative western blot (E) and quantitative data (F) of CD63 as an exosome marker in exosomes from TGF-β1- or GW4869-treated NRK-52E cells. Numbers (1 to 3) indicate each independent treatment in the given group. *p < 0.05 versus Ctrl-Exos, #p < 0.05 versus 5 ng/ml TGFβ1-Exos, &p < 0.05 versus 15 ng/ml TGFβ1-Exos (n = 3). (G) Fluorescent staining image of PKH-67-labeled NRK-52E cells. Scales bars=50 μm. (H) Fluorescent staining image of NRK-52E cell-derived exosomes taken up by NRK-49F cells. Scales bars=10 μm. (I, K) Representative western blot (I) and quantitative data (K) of α-SMA and PCNA in NRK-49F cells incubated with exosomes from TGF-β1- or GW4869-treated NRK-52E cells. Numbers (1 to 3) indicate each independent treatment in the giving group. *p < 0.05 versus Ctrl-Exos, #p < 0.05 versus 5 ng/ml TGFβ1-Exos, &p < 0.05 versus 15 ng/ml TGFβ1-Exos (n = 3). (J) Proliferation rate of NRK-49F cells incubated with NRK-52E cell-derived exosomes measured by CCK-8. *p < 0.05 versus Ctrl-Exos, #p < 0.05 versus 5 ng/ml TGFβ1-Exos, &p < 0.05 versus 15 ng/mL TGFβ1-Exos (n = 3). (L-N) Double immunofluorescence staining (green for Col-I and red for fibronectin) demonstrates the expression of Col-I and fibronectin in NRK-49F cells incubated with NRK-52E cell-derived exosomes. Scales bars=50 μm. *p < 0.05 versus Ctrl-Exos, #p < 0.05 versus 5 ng/ml TGFβ1-Exos, &p < 0.05 versus 15 ng/mL TGFβ1-Exos.
4). Small extracellular vesicles encapsulating lefty1 mRNA inhibit hepatic fibrosis. Asian Journal of Pharmaceutical Sciences (PubMed: 36382306) [IF=10.2]
5). Tubular cell-derived exosomal miR-150-5p contributes to renal fibrosis following unilateral ischemia-reperfusion injury by activating fibroblast in vitro and in vivo. International Journal of Biological Sciences (PubMed: 34671216) [IF=9.2]

Application: IF/ICC    Species: Mice    Sample: kidney

Figure 2 Increased secretion of exosomes in the kidney following UIRI. (A) Immunofluorescence staining for CD63 and TSG-101 (red). (B) Integrated optical density (IOD) of TSG-101. (C) Integrated optical density (IOD) of CD63. (D) Double immunofluorescence staining for CD63 (red) and AQP-1 (green), respectively. (E) Western blotting of CD63 in kidney tissue and exosome at different time points after UIRI (n=6). (F) Quantitative of CD63 in kidney. (G) Quantitative of CD63 in exosome. (H) TEM image of the exosomes in the kidney tissue after UIRI. *P < 0.05 versus sham.

Application: WB    Species: Mice    Sample: kidney

Figure 2 Increased secretion of exosomes in the kidney following UIRI. (A) Immunofluorescence staining for CD63 and TSG-101 (red). (B) Integrated optical density (IOD) of TSG-101. (C) Integrated optical density (IOD) of CD63. (D) Double immunofluorescence staining for CD63 (red) and AQP-1 (green), respectively. (E) Western blotting of CD63 in kidney tissue and exosome at different time points after UIRI (n=6). (F) Quantitative of CD63 in kidney. (G) Quantitative of CD63 in exosome. (H) TEM image of the exosomes in the kidney tissue after UIRI. *P < 0.05 versus sham.
6). MicroRNA-enriched small extracellular vesicles possess odonto-immunomodulatory properties for modulating the immune response of macrophages and promoting odontogenesis. Stem Cell Research & Therapy (PubMed: 33256846) [IF=7.5]

Application: WB    Species: Human    Sample: DPSCs-sEV

Fig. 1 Identification and characterization of DPSCs-sEV. a The morphology of DPSCs-sEV was determined by TEM, scale bar = 50 nm. b Expression of CD9 and CD63 in the DPSCs-sEV (sEV represents DPSCs-sEV and Lys represents DPSCs lysate, 尾-actin is a control for the lysate). c Nano-flow cytometry showed DPSCs-sEV ranged between 30 and 150 nm in diameter. d PKH26-labeled DPSCs-sEV were found in macrophage cytosol
7). Autophagy and Exosome Coordinately Enhance Macrophage M1 Polarization and Recruitment in Influenza A Virus Infection. Frontiers in Immunology (PubMed: 35371077) [IF=7.3]

Application: IF/ICC    Species: mouse    Sample: lung

FIGURE 3 | M1 polarization and LC3/CD63/IL-1b analysis of infected ANA-1 macrophages. Total RNA was isolated from infected ANA-1 cells at 24-h post-infection and used for transcriptional analysis of the related genes.(I) CD63 cellular immunofluorescence

Application: WB    Species: mouse    Sample: macrophages

FIGURE s4 |The total proteins of infected primary peritoneal macrophages were extracted at 24, 36, 48, and 72 h post-infection and then subjected for Western blotting. The corresponding antibodies were used to analyze autophagic protein (LC3, p62), exosome marker (CD63), and IL-1β activation pathway (IL-1β, cleaved IL-1β, and caspase-1) normalized to GAPDH.
8). Focal ischemic stroke modifies microglia-derived exosomal miRNAs: potential role of mir-212-5p in neuronal protection and functional recovery. Biological Research (PubMed: 37789455) [IF=6.7]

Application: WB    Species: Rat    Sample:

Fig. 2 MiRNA sequencing of microglial exosomes 3 days after MCAO/R and qRT–PCR validation. A, B Characterisation of microglial exosomes using nanoparticle tracking analysis and transmission electron microscopy scanning. Scale bar = 200 nm. C The exosome markers CD9, CD63, and CD81 were detected using western blot analysis. D Microglia-derived exosomes were detected by western blot analysis using the microglia marker CD11b. E Heatmap showing the levels of miRNAs in microglial exosomes at 3 days after MCAO/R. F Expression of miR-30c-5p, miR-126a-5p, miR-128-3p, miR-212-5p and miR-1949 in the ischemic penumbra of the cortex at 3 days after MCAO/R was determined using qRT–PCR. G Expression levels of the miRNA-targeting genes PLXNA2, PTEN and FOXO3 in the ischemic penumbra of the cortex at 3 days after MCAO/R. H The target sites of miR-212-5p in PLXNA2 mRNA 3’ untranslated region (3’UTR). I Map of the pmirGLO luciferase reporter vector. J Dual luciferase assays revealed the binding of miR-212-5p to the 3’UTR of PLXNA2. The data are presented as the means ± SEM (n = 5 per group). *P 
9). Chemerin-Induced Down-Regulation of Placenta-Derived Exosomal miR-140-3p and miR-574-3p Promotes Umbilical Vein Endothelial Cells Proliferation, Migration, and Tube Formation in Gestational Diabetes Mellitus. Cells (PubMed: 36359855) [IF=6.0]
10). OSCC cell-secreted exosomal CMTM6 induced M2-like macrophages polarization via ERK1/2 signaling pathway. Cancer immunology, immunotherapy : CII (PubMed: 33104837) [IF=5.8]
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