重组Anti-CD9抗体[EPR23105-125] (ab263019)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR23105-125] to CD9
- Suitable for: IP, WB, IHC-P, Flow Cyt
- Knockout validated
- Reacts with: Human
Related conjugates and formulations
概述
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产品名称
Anti-CD9抗体[EPR23105-125]
参阅全部 CD9 一抗 -
描述
兔单克隆抗体[EPR23105-125] to CD9 -
宿主
Rabbit -
经测试应用
适用于: IP, WB, IHC-P, Flow Cytmore details
不适用于: ICC/IF -
种属反应性
与反应: Human -
免疫原
Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
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阳性对照
- WB: HeLa, MCF7, HCT 116, HEL, Human tonsil and Human colon lysates. IHC-P: Human spleen and cervical carcinoma tissues. Flow Cyt: HCT 116 cells. IP: HCT 116 cell lysate.
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常规说明
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
性能
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形式
Liquid -
存放说明
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle. -
存储溶液
pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA -
Concentration information loading...
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纯度
Protein A purified -
克隆
单克隆 -
克隆编号
EPR23105-125 -
同种型
IgG -
研究领域
相关产品
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Alternative Versions
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Compatible Secondaries
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Corresponding Unmodified Peptide
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Isotype control
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KO cell lines
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KO cell lysates
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Recombinant Protein
应用
The Abpromise guarantee
Abpromise™承诺保证使用ab263019于以下的经测试应用
“应用说明”部分 下显示的仅为推荐的起始稀释度;实际最佳的稀释度/浓度应由使用者检定。
应用 | Ab评论 | 说明 |
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IP |
1/30.
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WB |
1/1000. Detects a band of approximately 22 kDa (predicted molecular weight: 25 kDa).
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IHC-P |
1/1000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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Flow Cyt |
1/500.
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说明 |
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IP
1/30. |
WB
1/1000. Detects a band of approximately 22 kDa (predicted molecular weight: 25 kDa). |
IHC-P
1/1000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
Flow Cyt
1/500. |
靶标
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功能
Involved in platelet activation and aggregation. Regulates paranodal junction formation. Involved in cell adhesion, cell motility and tumor metastasis. Required for sperm-egg fusion. -
组织特异性
Expressed by a variety of hematopoietic and epithelial cells. -
序列相似性
Belongs to the tetraspanin (TM4SF) family. -
翻译后修饰
Protein exists in three forms with molecular masses between 22 and 27 kDa, and is known to carry covalently linked fatty acids. -
细胞定位
Membrane. - Information by UniProt
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数据库链接
- Entrez Gene: 928 Human
- Omim: 143030 Human
- SwissProt: P21926 Human
- Unigene: 114286 Human
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别名
- Tetraspanin 29 antibody
- 5H9 antibody
- 5H9 antigen antibody
see all
图片
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All lanes : Anti-CD9 antibody [EPR23105-125] (ab263019) at 1/1000 dilution
Lane 1 : Wild-type HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate
Lane 2 : CD9 knockout HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate
Lane 3 : A549 (Human lung carcinoma cell line) whole cell lysate
Lane 4 : MCF7 (Human breast adenocarcinoma cell line) whole cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 25 kDa
Observed band size: 18 kDa why is the actual band size different from the predicted?Lanes 1 - 4: Merged signal (red and green). Green - ab263019 observed at 18 kDa. Red - loading control ab7291 (Mouse anti-Alpha Tubulin [DM1A]) observed at 55 kDa.
ab263019 was shown to react with CD9 in wild-type HeLa cells in Western blot with loss of signal observed in CD9 knockout cell line ab255375 (CD9 knockout cell lysate ab263754). Wild-type HeLa and CD9 knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in fluorescent western blot (TBS-based) blocking solution before incubation with ab263019 and ab7291 (Mouse anti-Alpha Tubulin [DM1A]) overnight at 4 °C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 h at room temperature before imaging.
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Immunohistochemical analysis of paraffin-embedded Human cervical carcinoma tissue labeling CD9 with ab263019 at 1/1000 dilution followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on human cervical carcinoma. The section was incubated with ab263019 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).
Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20mins.
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Flow cytometric analysis of Raji (human Burkitt's lymphoma B lymphocyte, Left) / HCT 116 (human colorectal carcinoma epithelial cell, Right) cells labelling CD9 with ab263019 at 1/500 dilution compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) at 1/2000 was used as the secondary antibody.
Negative control: Raji (PMID: 8921952).
Gated on viable cells.
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All lanes : Anti-CD9 antibody [EPR23105-125] (ab263019) at 1/1000 dilution
Lane 1 : HEL (erythroleukemia erythroblast), whole cell lysate
Lane 2 : Human tonsil lysate
Lane 3 : Human colon lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Predicted band size: 25 kDa
Observed band size: 22 kDa why is the actual band size different from the predicted?Blocking and diluting buffer and concentration: 5% NFDM/TBST.
The molecular weight observed is consistent with what has been described in the literature (PMID: 3275469). Exposure time: 3 minutes.
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All lanes : Anti-CD9 antibody [EPR23105-125] (ab263019) at 1/1000 dilution
Lane 1 : HCT 116 (human colorectal carcinoma epithelial cell) whole cell lysate
Lane 2 : Raji (human Burkitts lymphoma B lymphocyte) whole cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Predicted band size: 25 kDa
Observed band size: 22 kDa why is the actual band size different from the predicted?Blocking and diluting buffer and concentration: 5% NFDM/TBST.
The molecular weight observed is consistent with what has been described in the literature (PMID: 3275469).
Negative control: Raji (PMID: 8921952). Exposure time: 3 minutes.
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CD9 was immunoprecipitated from 0.35 mg HCT 116 (human colorectal carcinoma epithelial cell) whole cell lysate with ab263019 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab263019 at 1/500 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366) was used at 1/1000 dilution.
Lane 1: HCT 116 (human colorectal carcinoma epithelial cell) whole cell lysate 10ug.
Lane 2: ab263019 IP in HCT 116 whole cell lysate.
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab263019 in HCT 116 whole cell lysate.
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 10 seconds.
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Immunohistochemical analysis of paraffin-embedded Human spleen tissue labeling CD9 with ab263019 at 1/1000 dilution followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on platelets of human spleen. The section was incubated with ab263019 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).
Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.
实验方案
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
数据表及文件
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SDS download
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Datasheet download
Certificate of Compliance
文献 (10)
ab263019 被引用在 10 文献中.
- Li X et al. Eugenol-Preconditioned Mesenchymal Stem Cell-Derived Extracellular Vesicles Promote Antioxidant Capacity of Tendon Stem Cells In Vitro and In Vivo. Oxid Med Cell Longev 2022:3945195 (2022). PubMed: 35178155
- Hu K et al. Presentation of antigen on extracellular vesicles using transmembrane domains from viral glycoproteins for enhanced immunogenicity. J Extracell Vesicles 11:e12199 (2022). PubMed: 35233930
- Xu W et al. Plasma-Derived Exosomal SncRNA as a Promising Diagnostic Biomarker for Early Detection of HBV-Related Acute-on-Chronic Liver Failure. Front Cell Infect Microbiol 12:923300 (2022). PubMed: 35873157
- Huang X et al. La protein regulates protein expression by binding with the mRNAs of target genes and participates the pathological process of ovarian cancer. Front Oncol 12:763480 (2022). PubMed: 36110943
- Malkin EZ et al. Cell-free DNA topology depends on its subcellular and cellular origins in cancer. JCI Insight 7:N/A (2022). PubMed: 36125881
- Zhu G et al. LncRNA XIST from the bone marrow mesenchymal stem cell derived exosome promotes osteosarcoma growth and metastasis through miR-655/ACLY signal. Cancer Cell Int 22:330 (2022). PubMed: 36309693
- Li H et al. Knockdown of circ-FANCA alleviates LPS-induced HK2 cell injury via targeting miR-93-5p/OXSR1 axis in septic acute kidney injury. Diabetol Metab Syndr 13:7 (2021). PubMed: 33468219
- Zhang S et al. Exosomal miR-183-5p Shuttled by M2 Polarized Tumor-Associated Macrophage Promotes the Development of Colon Cancer via Targeting THEM4 Mediated PI3K/AKT and NF-κB Pathways. Front Oncol 11:672684 (2021). PubMed: 34249713
- Huang Z et al. Weighted gene co-expression network analysis identifies key genes from extracellular vesicles as potential prognostic biomarkers for congenital pulmonary stenosis. Mol Med Rep 22:2528-2536 (2020). PubMed: 32705286
- Chen X et al. Exosomal Long Non-coding RNA HOTTIP Increases Resistance of Colorectal Cancer Cells to Mitomycin via Impairing MiR-214-Mediated Degradation of KPNA3. Front Cell Dev Biol 8:582723 (2020). PubMed: 33585440