CD9 Antibody - #AF5139

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Product: CD9 Antibody
Catalog: AF5139
Description: Rabbit polyclonal antibody to CD9
Application: WB IHC IF/ICC
Reactivity: Human, Mouse, Rat
Prediction: Pig, Bovine, Horse, Sheep, Rabbit, Dog
Mol.Wt.: 20~35kD; 25kD(Calculated).
Uniprot: P21926
RRID: AB_2837625

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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:
WB 1:500-1:2000, IHC 1:50-1:200, IF/ICC 1:100-1:500
*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
Prediction:
Pig(82%), Bovine(91%), Horse(100%), Sheep(91%), Rabbit(91%), Dog(82%)
Clonality:
Polyclonal
Specificity:
CD9 Antibody detects endogenous levels of total CD9.
RRID:
AB_2837625
Cite Format: Affinity Biosciences Cat# AF5139, RRID:AB_2837625.
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

Tetraspanin 29; 5H9; 5H9 antigen; Antigen defined by monoclonal antibody 602 29; Antigen defined by monoclonal antibody 60229; BA-2/p24 antigen; BA2; BTCC 1; BTCC1; CD9; CD9 antigen; CD9 antigen p24; CD9 molecule; CD9_HUMAN; Cell growth inhibiting gene 2 protein; Cell growth-inhibiting gene 2 protein; DRAP 27; DRAP27; GIG2; Growth inhibiting gene 2 protein; Leukocyte antigen MIC3; MIC3; Motility related protein; Motility-related protein; MRP 1; MRP-1; MRP1; p24; p24 antigen; Tetraspanin-29; Tspan 29; Tspan-29; TSPAN29;

Immunogens

Immunogen:
Uniprot:

P21926(CD9_HUMAN) >>Visit HPA database.

Gene(ID):
CD9(928)
Expression:
P21926 CD9_HUMAN:

Detected in platelets (at protein level) (PubMed:19640571). Expressed by a variety of hematopoietic and epithelial cells (PubMed:19640571).

Description:
Involved in platelet activation and aggregation. Regulates paranodal junction formation. Involved in cell adhesion, cell motility and tumor metastasis. Required for sperm-egg fusion.
Sequence:
MPVKGGTKCIKYLLFGFNFIFWLAGIAVLAIGLWLRFDSQTKSIFEQETNNNNSSFYTGVYILIGAGALMMLVGFLGCCGAVQESQCMLGLFFGFLLVIFAIEIAAAIWGYSHKDEVIKEVQEFYKDTYNKLKTKDEPQRETLKAIHYALNCCGLAGGVEQFISDICPKKDVLETFTVKSCPDAIKEVFDNKFHIIGAVGIGIAVVMIFGMIFSMILCCAIRRNREMV

Predictions

Predictions:

Score>80(red) has high confidence and is suggested to be used for WB detection. *The prediction model is mainly based on the alignment of immunogen sequences, the results are for reference only, not as the basis of quality assurance.

Species
Results
Score
Horse
100
Bovine
91
Sheep
91
Rabbit
91
Pig
82
Dog
82
Xenopus
50
Zebrafish
0
Chicken
0
Model Confidence:
High(score>80) Medium(80>score>50) Low(score<50) No confidence

PTMs - P21926 As Substrate

Site PTM Type Enzyme
Y125 Phosphorylation
K126 Methylation
K126 Ubiquitination
K131 Ubiquitination
K133 Ubiquitination
K135 Ubiquitination
K170 Ubiquitination
T175 Phosphorylation
T177 Phosphorylation
K179 Ubiquitination
K186 Ubiquitination

Research Backgrounds

Function:

Integral membrane protein associated with integrins, which regulates different processes, such as sperm-egg fusion, platelet activation and aggregation, and cell adhesion. Present at the cell surface of oocytes and plays a key role in sperm-egg fusion, possibly by organizing multiprotein complexes and the morphology of the membrane required for the fusion (By similarity). In myoblasts, associates with CD81 and PTGFRN and inhibits myotube fusion during muscle regeneration (By similarity). In macrophages, associates with CD81 and beta-1 and beta-2 integrins, and prevents macrophage fusion into multinucleated giant cells specialized in ingesting complement-opsonized large particles. Also prevents the fusion between mononuclear cell progenitors into osteoclasts in charge of bone resorption (By similarity). Acts as a receptor for PSG17 (By similarity). Involved in platelet activation and aggregation. Regulates paranodal junction formation (By similarity). Involved in cell adhesion, cell motility and tumor metastasis.

PTMs:

Palmitoylated at a low, basal level in unstimulated platelets. The level of palmitoylation increases when platelets are activated by thrombin (in vitro). The protein exists in three forms with molecular masses between 22 and 27 kDa, and is known to carry covalently linked fatty acids.

Subcellular Location:

Cell membrane>Multi-pass membrane protein. Membrane>Multi-pass membrane protein. Secreted>Extracellular exosome.
Note: Present at the cell surface of oocytes. Accumulates in the adhesion area between the sperm and egg following interaction between IZUMO1 and its receptor IZUMO1R/JUNO.

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). Expressed by a variety of hematopoietic and epithelial cells.

Subunit Structure:

Forms both disulfide-linked homodimers and higher homooligomers as well as heterooligomers with other members of the tetraspanin family. Interacts (via the second extracellular domain) with integrin ITGAV:ITGB3. Interacts with integrin ITGA6:ITGB1; interaction takes place in oocytes and is involved in sperm-egg fusion (By similarity). Part of integrin-tetraspanin complexes composed of CD81, beta-1 and beta-2 integrins in the membrane of monocyte/macrophages. Interacts with CD63; identified in a complex with CD63 and ITGB3. Associates with CR2/CD21 and with PTGFRN/CD9P1. Part of a complex composed of CD9, CD81, PTGFRN and IGSF8 (By similarity). Interacts directly with IGSF8. Interacts with PDPN; this interaction is homophilic and attenuates platelet aggregation and pulmonary metastasis induced by PDPN. Interacts (on T cell side) with CD81 at immunological synapses between antigen-presenting cells and T cells.

Family&Domains:

Belongs to the tetraspanin (TM4SF) family.

Research Fields

· Organismal Systems > Immune system > Hematopoietic cell lineage.   (View pathway)

References

1). Iron Oxide Nanoparticles Engineered Macrophage-Derived Exosomes for Targeted Pathological Angiogenesis Therapy. ACS nano, 2024 (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.
2). Exosome-mediated delivery of inflammation-responsive Il-10 mRNA for controlled atherosclerosis treatment. Theranostics, 2023 (PubMed: 34815799) [IF=12.4]

Application: WB    Species: Human    Sample: HEK293T cells

Figure 2 Preparation and characterization of ExoIRES-IL-10. (A) Schematic of the exosomes preparation and isolation process. (B) Western blot analysis of the exosomal inclusive markers (TSG101, CD9), exclusive marker (GM130), negative markers to confirm the purity (APOA1) in the isolated exosomes and parental cells. HEK293T cells were treated with PBS or transfected, empty vector, or IRES-Il-10 plasmids. (C) Representative transmission electron microscope (TEM) images of ExoNone, ExoEmpty or ExoIRES-IL-10. Scale bar = 200 nm. (D) Size distribution of the isolated exosomes as indicated. (E) qPCR analysis of Il-10 mRNA in the isolated exosomes as indicated. GAPDH served as an internal control. (F) qPCR analysis of Il-10 mRNA in ExoIRES-IL-10 in exosomal RNA degradation assay as indicated. GAPDH served as an internal control. Data are presented as mean ± SEM of three independent experiments. ND, not determined as Ct value greater than 38.
3). Isothiazolinone dysregulates the pattern of miRNA secretion: Endocrine implications for neurogenesis. Environment international, 2023 (PubMed: 37939439) [IF=11.8]
4). Chondrocyte-targeted exosome-mediated delivery of Nrf2 alleviates cartilaginous endplate degeneration by modulating mitochondrial fission. Journal of nanobiotechnology, 2024 (PubMed: 38790015) [IF=10.2]

Application: WB    Species: Human    Sample: HEK293T cells

Fig. 1 Construction and features of engineered CAP-Nrf2-Exos. (A, B) Western blot and qRT‒PCR analysis showing the mRNA and protein expression levels of Nrf2 in HEK293T cells. (C, D) Western blot and qRT‒PCR analysis of Nrf2 in the engineered exosomes. (E, F) Western blot and qRT‒PCR analysis of Nrf2 expression in CEP cells after treatment with engineered exosomes. (G) Western blot analysis of Lamp2b, CD9, CD63, TSG101, and β-Actin in HEK293T cells, HEK293T cells transfected with CAP-Nrf2 overexpression plasmids, Exos and CAP-Nrf2-Exos. (H) Exos and CAP-Nrf2-Exos were examined by NTA. (I) TEM analysis of Exos and CAP-Nrf2-Exos isolated from the culture medium of HEK293T cells. Scale bar, 100 nm. (J) Fluorescence images of PKH26-labelled Exos and CAP-Exos internalized by CEP cells. Scale bar, 50 μm and 10 μm. (K) Fluorescence images of DiR-labelled Exos and CAP-Exos in the CEP and IVD in the groups mentioned above. (n = 3, *p 
5). Extracellular vesicles delivering nuclear factor I/C for hard tissue engineering: Treatment of apical periodontitis and dentin regeneration. Journal of Tissue Engineering, 2022 (PubMed: 35321254) [IF=8.2]

Application: WB    Species: Human    Sample: SCAPs

Figure 2. Identification of EVs derived from LPS-stimulated DPCs (LPS-EVs) and establishment of in vitro model. (a) CCK8 assay detected cell viability of DPCs treated LPS (0.1, 1, 10, 100 µg/mL). (b) 1 µg/mL LPS increased EVs secretion of DPCs, compared to the other groups. (c) Schematic diagram shows the four types EV collected from EV-free culture medium of DPCs with or without LPS stimulation (0.1, 1, 10 µg/mL), which were used in subsequent assays. (d) Nanoparticle tracing assay (NTA) revealed the diameter of collected EVs was approximately 100 nm. (e) Scanning electron microscopy (SEM) of cup and saucer-shaped EVs. (f) Immunofluorescence staining confirmed that PKH26-labeled EVs (red) were endocytosed by SCAPs. DAPI (blue), and F-actin (green). (g) Western blot analysis of EVs surface markers (CD9, CD63, CD81 and TSG101). (h) CCK8 assay detected the effect of Nor-EV and LPS-EVs on cell viability of SCAPs. *p < 0.05. **p < 0.01. ***p < 0.001.#p < 0.0001.
6). MicroRNA-enriched small extracellular vesicles possess odonto-immunomodulatory properties for modulating the immune response of macrophages and promoting odontogenesis. Stem Cell Research & Therapy, 2020 (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). Endometrial extracellular vesicles from women with recurrent implantation failure attenuate the growth and invasion of embryos. FERTILITY AND STERILITY, 2020 (PubMed: 32622655) [IF=6.7]

Application: WB    Species: Human    Sample: RIF-EVs and FER-EVs.

Figure 1.Isolation of EVs and determination of RIF-EVs and FER-EVs. (A) Western blotting shows that RIF-EVs and FER-EVs expressed classic EV protein markers Alix, TSG101, and CD9. Representative shapes of (B) RIF-EVs and (C) FER-EVs detected with the use of transmission electron microscopy. The size and distribution of (D) RIF-EVs and (E) FER-EVs examined with the use of nanoparticle tracking analysis. ECs ¼ endometrial cells; EVs ¼ extracellular vesicles; FER ¼ fertile women; RIF ¼ women with recurrent implantation failure. Liu. Extracellular vesicles regulate embryos. Fertil Steril 2020.
8). Mesenchymal stem cell-derived exosome mitigates colitis via the modulation of the gut metagenomics–metabolomics–farnesoid X receptor axis. Biomaterials Science, 2022 (PubMed: 35858469) [IF=6.6]
9). CD9 exacerbates pathological cardiac hypertrophy through regulating GP130/STAT3 signaling pathway. iScience, 2023 (PubMed: 37860696) [IF=5.8]
10). CD73-Positive Small Extracellular Vesicles Derived From Umbilical Cord Mesenchymal Stem Cells Promote the Proliferation and Migration of Pediatric Urethral Smooth Muscle Cells Through Adenosine Pathway. Frontiers in Bioengineering and Biotechnology, 2022 (PubMed: 35573239) [IF=5.7]

Application: WB    Species: Human    Sample: PUSMCs

FIGURE 1 Identification of PUMSCs, UCMSCs and UCMSC-sEV. (A) Morphology of PUMSCs. (B) Immunofluorescence staining results show that the isolated PUMSCs express α-SMA, a smooth muscle cell surface marker. (C) Growth curve of PUMSCs. (D) Flow cytometric analysis of MSC surface markers shows that UCMSCs express high levels = of CD29, CD44, CD73, and CD90 but do not express CD45 and HLA-DR. (E) Transmission electron microscopy shows that the UCMSC-sEV are cup-shaped vesicles. Scale bar = 100 nm. (F) NanoFCM analysis of the particle size of UCMSC-sEV. (G) Western blotting was used to detect the expression of the sEV marker proteins CD9, CD63, CD81, and TSG101.
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