重组Anti-CD9抗体[EPR27551-92] (ab307085)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR27551-92] to CD9
- Suitable for: IHC-P, WB, IP
- Knockout validated
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
概述
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产品名称
Anti-CD9抗体[EPR27551-92]
参阅全部 CD9 一抗 -
描述
兔单克隆抗体[EPR27551-92] to CD9 -
宿主
Rabbit -
特异性
Unsuitable for human IHC-P.
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经测试应用
适用于: IHC-P, WB, IPmore details
不适用于: Flow Cyt,ICC/IF or IHC-Fr -
种属反应性
与反应: Mouse, Rat, 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: Wild-type HeLa, CD9 knockout HeLa whole cell lysate, MCF7, RAW 264.7, HCT 116, K-562, Raji, Mouse brain, Mouse kidney and C6 lysates. IHC-P: Mouse cerebrum, Mouse kidney, Mouse lung cancer, Rat cerebrum and Rat kidney tissues. IP: RAW 264.7 whole 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. Avoid freeze / thaw cycle. -
存储溶液
pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 40% Glycerol (glycerin, glycerine), 0.05% BSA, 59% PBS -
Concentration information loading...
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纯度
Protein A purified -
克隆
单克隆 -
克隆编号
EPR27551-92 -
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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Isotype control
应用
The Abpromise guarantee
Abpromise™承诺保证使用ab307085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 |
---|---|---|
IHC-P |
1/2000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
This antibody is not suitable for human species in IHC-P application. |
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WB |
1/1000. Detects a band of approximately 23 kDa.
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IP |
1/30.
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说明 |
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IHC-P
1/2000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. This antibody is not suitable for human species in IHC-P application. |
WB
1/1000. Detects a band of approximately 23 kDa. |
IP
1/30. |
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
- Entrez Gene: 12527 Mouse
- Entrez Gene: 24936 Rat
- Omim: 143030 Human
- SwissProt: P21926 Human
- SwissProt: P40240 Mouse
- SwissProt: P40241 Rat
- Unigene: 114286 Human
see all -
别名
- Tetraspanin 29 antibody
- 5H9 antibody
- 5H9 antigen antibody
see all
图片
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All lanes : Anti-CD9 antibody [EPR27551-92] (ab307085) at 1/1000 dilution
Lane 1 : Wild-type HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate 20 µg
Lane 2 : CD9 knockout HeLa whole cell lysate 20 µg
Lane 3 : MCF7 (human breast adenocarcinoma epithelial cell) whole cell lysate 20 µg
Lane 4 : RAW 264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage) whole cell lysate 20 µg
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (IRDye® 800CW) (ab216773) and Goat Anti-Mouse IgG H&L (IRDye® 680RD) (ab216776) at 1/10000 dilution
Observed band size: 23 kDa why is the actual band size different from the predicted?Blocking and diluting buffer and concentration: Intercept® (TBS) Blocking Buffer diluted with an equal volume of 0.1% TBST
The samples were run on a Bis-Tris gel.
Performed under reducing conditions.
False colour image of Western blot: Anti-CD9 antibody [EPR27551-92] (ab307085) staining at 1/1000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution, shown in red.
In Western blot, ab307085 was shown to bind specifically to CD9. A band was observed at 23 kDa in wild-type HeLa cell lysates whereas no signal observed at this size in CD9 knockout cell line ab255375 (knockout cell lysate ab263754). To generate this image, wild-type and CD9 knockout HeLa cell lysates were analyzed. First, samples were run on an SDS-PAGE gel then transferred onto an immobilon-FL PVDF membrane. Membranes were blocked in in Intercept® (TBS) Blocking Buffer diluted with an equal volume of 0.1% TBS 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 (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) at 1/20000 dilution.
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All lanes : Anti-CD9 antibody [EPR27551-92] (ab307085) at 1/1000 dilution
Lane 1 : HCT 116 (human colorectal carcinoma epithelial cell) whole cell lysate 20 µg
Lane 2 : K-562 (human chronic myelogenous leukemia lymphoblast) whole cell lysate 20 µg
Lane 3 : Raji (human Burkitts lymphoma B lymphocyte) whole cell lysate 20 µg
Lane 4 : Mouse brain tissue lysate 20 µg
Lane 5 : Mouse kidney tissue lysate 20 µg
Lane 6 : C6 (rat glial tumor glial cell) whole cell lysate 20 µg
Secondary
All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1/100000 dilution
Observed band size: 23 kDa why is the actual band size different from the predicted?Blocking and diluting buffer and concentration: 5% NFDM/TBST
Negative controls: K-562, Raji (PMID:17407154 ; PMID: 8921952).
Exposure time: 70 seconds
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CD9 was immunoprecipitated from 0.35 mg RAW 264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage) whole cell lysate 10 ug with ab307085 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab307085 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution.
Lane 1: RAW 264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage) whole cell lysate 10 ug
Lane 2: abab307085 IP in RAW 264.7 whole cell lysate
Lane 3:Rabbit monoclonal IgG (ab172730) instead of ab307085 in RAW 264.7 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 Rat kidney tissue labeling CD9 with ab307085 at 1/2000 (0.228 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) was used. Membranous staining in rat kidney. The section was incubated with ab307085 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 LeicaDS9800 (Bond™ Polymer Refine Detection) was used.
Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins
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Immunohistochemical analysis of paraffin-embedded Rat cerebrum tissue labeling CD9 with ab307085 at 1/2000 (0.228 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) was used. Positive staining in rat cerebrum. The section was incubated with ab307085 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 LeicaDS9800 (Bond™ Polymer Refine Detection) was used.
Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins
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Immunohistochemical analysis of paraffin-embedded Mouse lung cancer tissue labeling CD9 with ab307085 at 1/2000 (0.228 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) was used. Membranous staining in mouse lung cancer. The section was incubated with ab307085 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 LeicaDS9800 (Bond™ Polymer Refine Detection) was used.
Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins
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Immunohistochemical analysis of paraffin-embedded Mouse kidney tissue labeling CD9 with ab307085 at 1/2000 (0.228 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) was used. Positive staining in mouse kidney. The section was incubated with ab307085 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 LeicaDS9800 (Bond™ Polymer Refine Detection) was used.
Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins
-
Immunohistochemical analysis of paraffin-embedded Mouse cerebrum tissue labeling CD9 with ab307085 at 1/2000 (0.228 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) was used. Positive staining in mouse cerebrum. The section was incubated with ab307085 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 LeicaDS9800 (Bond™ Polymer Refine Detection) was used.
Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) 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 (1)
ab307085 被引用在 1 文献中.
- Sun J et al. Engineered small extracellular vesicles loaded with miR-654-5p promote ferroptosis by targeting HSPB1 to alleviate sorafenib resistance in hepatocellular carcinoma. Cell Death Discov 9:362 (2023). PubMed: 37777559