重组Anti-ICAM1抗体[EPR24639-3] (ab282575)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR24639-3] to ICAM1
- Suitable for: Flow Cyt, IP, IHC-P, ICC/IF, WB
- Knockout validated
- Reacts with: Rat, Human
Related conjugates and formulations
概述
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产品名称
Anti-ICAM1抗体[EPR24639-3]
参阅全部 ICAM1 一抗 -
描述
兔单克隆抗体[EPR24639-3] to ICAM1 -
宿主
Rabbit -
特异性
Rat species is recommended based on Flow Cyt result. We do not guarantee other applications for rat.
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经测试应用
适用于: Flow Cyt, IP, IHC-P, ICC/IF, WBmore details
不适用于: IHC-Fr -
种属反应性
与反应: 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 whole cell lysate. Ramos and Raji whole cell lysate. Human kidney tissue lysate. IHC-P: Human kidney tissue. Human breast cancer tissue. ICC/IF: Raji cells. Wild-type HeLa cells. Flow Cyt: Rat splenocytes. C6 and Raji cells. IP: Raji 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: 7.2
Preservative: 0.01% Sodium azide
Constituents: 59.94% 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
EPR24639-3 -
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
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KO cell lines
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KO cell lysates
应用
The Abpromise guarantee
Abpromise™承诺保证使用ab282575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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Flow Cyt |
1/500.
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IP |
1/30.
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IHC-P |
1/600. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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ICC/IF |
1/500.
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WB | (1) |
1/1000. Predicted molecular weight: 57 kDa.
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说明 |
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Flow Cyt
1/500. |
IP
1/30. |
IHC-P
1/600. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
ICC/IF
1/500. |
WB
1/1000. Predicted molecular weight: 57 kDa. |
靶标
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功能
ICAM proteins are ligands for the leukocyte adhesion protein LFA-1 (integrin alpha-L/beta-2). During leukocyte trans-endothelial migration, ICAM1 engagement promotes the assembly of endothelial apical cups through ARHGEF26/SGEF and RHOG activation. In case of rhinovirus infection acts as a cellular receptor for the virus. -
序列相似性
Belongs to the immunoglobulin superfamily. ICAM family.
Contains 5 Ig-like C2-type (immunoglobulin-like) domains. -
翻译后修饰
Monoubiquitinated, which is promoted by MARCH9 and leads to endocytosis. -
细胞定位
Membrane. - Information by UniProt
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数据库链接
- Entrez Gene: 3383 Human
- Entrez Gene: 25464 Rat
- Omim: 147840 Human
- SwissProt: P05362 Human
- SwissProt: Q00238 Rat
- Unigene: 643447 Human
- Unigene: 12 Rat
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别名
- Antigen identified by monoclonal antibody BB2 antibody
- BB 2 antibody
- BB2 antibody
see all
图片
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All lanes : Anti-ICAM1 antibody [EPR24639-3] (ab282575) at 1/1000 dilution
Lane 1 : Wild-type HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate
Lane 2 : ICAM1 knockout HeLa whole cell lysate
Lane 3 : Ramos (Human Burkitt's lymphoma B lymphocyte) whole cell lysate
Lysates/proteins at 20 µg per lane.
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
Predicted band size: 57 kDa
Observed band size: 90 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% TBS
Lanes 1-3: Merged signal (red and green). Green - ab282575 observed at 90kDa. Red - loading control ab8245 observed at 36 kDa.
ab282575 Anti-ICAM1 antibody [EPR24639-3] was shown to react with ICAM1 in wild-type Hela cells in Western blot. Loss of signal was observed when knockout cell line ab261742 (knockout cell lysate ab256947) was used. Wild-type and ICAM1 knockout samples were subjected to SDS-PAGE.
ab282575 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated at 4°C overnight at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 10000 dilution for 1 hour at room temperature before imaging.
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All lanes : Anti-ICAM1 antibody [EPR24639-3] (ab282575) at 1/1000 dilution
Lane 1 : Wild-type HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate
Lane 2 : ICAM1 knockout Hela whole cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 57 kDa
Observed band size: 90 kDa why is the actual band size different from the predicted?Blocking and diluting buffer and concentration: 5% NFDM/TBST
ab282575 Anti-ICAM1 antibody [EPR24639-3] was shown to react with ICAM1 in wild-type Hela cells in Western blot. Loss of signal was observed when knockout cell line ab261742 (knockout cell lysate ab256947) was used. Wild-type and ICAM1 knockout samples were subjected to SDS-PAGE.
Exposure time: 15 seconds.
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Anti-ICAM1 antibody [EPR24639-3] (ab282575) at 1/1000 dilution + human kidney tissue lysate at 20 µg
Secondary
VeriBlot for IP Detection Reagent (HRP) (ab131366) at 1/1000 dilution
Predicted band size: 57 kDa
Observed band size: 90 kDa why is the actual band size different from the predicted?Blocking and diluting buffer and concentration: 5% NFDM/TBST
Exposure time: 26 seconds
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All lanes : Anti-ICAM1 antibody [EPR24639-3] (ab282575) at 1/1000 dilution
Lane 1 : Ramos (human Burkitt's lymphoma B lymphocyte) 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 1/20000 dilution
Predicted band size: 57 kDa
Observed band size: 90 kDa why is the actual band size different from the predicted?Blocking and diluting buffer and concentration: 5% NFDM/TBST
Exposure time: 15 seconds
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Immunohistochemical analysis of paraffin-embedded human kidney tissue labelling ICAM1 with ab282575 at 1/600 (0.902 µg/ml) dilution followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on normal human glomerulus. The section was incubated with ab282575 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).
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 human breast cancer tissue labelling ICAM1 with ab282575 at 1/600 (0.902 µg/ml) dilution followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on human breast cancer (PMID: 30082828). The section was incubated with ab282575 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).
Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.
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Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized Raji cells labelling ICAM1 with ab282575 at 1/500 (1.082 µg/ml) dilution, followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) antibody at 1/1000 dilution (Green). Confocal image showing membranous staining in Raji cells.
ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 dilution (Red). The nuclear counterstain was DAPI (Blue).
Secondary antibody only control: Secondary antibody is ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) at 1/1000 dilution.
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Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized ICAM1 KO HeLa cells labelling ICAM1 with ab282575 at 1/500 (1.082 µg/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 dilution (Green). Confocal image showing membranous staining in wild-type HeLa cells, and no staining in ICAM1 knockout HeLa cells is observed.
ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 dilution (Red). The nuclear counterstain was DAPI (Blue).
Secondary antibody only control: Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution.
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Flow cytometric analysis of rat splenocytes cells labelling ICAM1 with ab282575 at 1/500 dilution (0.1 µg)/ Right compared with a Rabbit monoclonal IgG (ab172730) / Left isotype control. A goat anti rabbit IgG (Alexa Fluor® 488, ab150077) at 1/2000 dilution was used as the secondary antibody.
Gated on viable cells.
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Flow cytometric analysis of C6 (Rat glial tumor glial cell) cells labelling ICAM1 with ab282575 at 1/500 dilution (0.1 µg)/(red) 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 dilution was used as the secondary antibody.
Gated on viable cells.
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Flow cytometric analysis of Raji (Human Burkitt's lymphoma B lymphocyte) cells labelling ICAM1 with ab282575 at 1/500 dilution (0.1 µg)/(Red) 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 dilution was used as the secondary antibody.
Gated on viable cells.
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ICAM1 was immunoprecipitated from 0.35 mg Raji (human burkitt's lymphoma b lymphocyte) whole cell lysate 10 µg with ab282575 at 1/30 dilution (2 µg in 0.35 mg lysates). Western blot was performed on the immunoprecipitate using ab282575 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution.
Lane 1: Raji whole cell lysate 10 µg
Lane 2: ab282575 IP in Raji whole cell lysate
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab282575 in Raji whole cell lysate
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 3 minutes.
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 (4)
ab282575 被引用在 4 文献中.
- Chen Y et al. Taxifolin improves inflammatory injury of human bronchial epithelial cells by inhibiting matrix metalloproteinase (MMP) 10 via Wnt/β-catenin pathway. Bioengineered 13:1198-1208 (2022). PubMed: 35000533
- Tan Z et al. KLF2 alleviates endothelial cell injury and inhibits the formation of THP‑1 macrophage‑derived foam cells by activating Nrf2 and enhancing autophagy. Exp Ther Med 24:737 (2022). PubMed: 36478888
- Huang HW et al. Fibroblast Growth Factor Type 1 Ameliorates High-Glucose-Induced Oxidative Stress and Neuroinflammation in Retinal Pigment Epithelial Cells and a Streptozotocin-Induced Diabetic Rat Model. Int J Mol Sci 22:N/A (2021). PubMed: 34281287
- Yan Z et al. Cancer-associated fibroblast-derived exosomal miR-18b promotes breast cancer invasion and metastasis by regulating TCEAL7. Cell Death Dis 12:1120 (2021). PubMed: 34853307