重组Anti-ICAM1抗体[EP1442Y] - Low endotoxin,Azide free (ab221777)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EP1442Y] to ICAM1 - Low endotoxin, Azide free
- Suitable for: WB, IHC-P
- Reacts with: Human
Related conjugates and formulations
概述
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产品名称
Anti-ICAM1抗体[EP1442Y] - Low endotoxin,Azide free
参阅全部 ICAM1 一抗 -
描述
兔单克隆抗体[EP1442Y] to ICAM1 - Low endotoxin,Azide free -
宿主
Rabbit -
经测试应用
适用于: WB, IHC-Pmore details -
种属反应性
与反应: Human -
免疫原
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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阳性对照
- WB: Huvec, Ramos and Raji whole cell lysate; IHC: Human kidney and tonsil tissue.
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常规说明
ab221777 is the carrier-free version of ab53013.
Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.
This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.
Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.
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.
Our Low endotoxin, azide-free formats have low endotoxin level (≤ 1 EU/ml, determined by the LAL assay) and are free from azide, to achieve consistent experimental results in functional assays.
Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.
性能
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形式
Liquid -
存放说明
Shipped at 4°C. Store at +4°C. Do Not Freeze. -
存储溶液
pH: 7.20
Constituent: PBS -
无载体
是 -
Concentration information loading...
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纯度
Protein A purified -
克隆
单克隆 -
克隆编号
EP1442Y -
同种型
IgG -
研究领域
相关产品
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Alternative Versions
- Anti-ICAM1 antibody [EP1442Y] - BSA and Azide free (ab271852)
- Alexa Fluor® 488 Anti-ICAM1 antibody [EP1442Y] (ab311054)
- Alexa Fluor® 647 Anti-ICAM1 antibody [EP1442Y] (ab311180)
- Alexa Fluor® 594 Anti-ICAM1 antibody [EP1442Y] (ab311835)
- Alexa Fluor® 568 Anti-ICAM1 antibody [EP1442Y] (ab313118)
- Alexa Fluor® 555 Anti-ICAM1 antibody [EP1442Y] (ab313311)
- Anti-ICAM1 antibody [EP1442Y] (ab53013)
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Compatible Secondaries
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Conjugation kits
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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ab221777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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WB |
Use at an assay dependent concentration. Detects a band of approximately 89 kDa (predicted molecular weight: 58 kDa).
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IHC-P |
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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说明 |
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WB
Use at an assay dependent concentration. Detects a band of approximately 89 kDa (predicted molecular weight: 58 kDa). |
IHC-P
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
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
- Omim: 147840 Human
- SwissProt: P05362 Human
- Unigene: 643447 Human
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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 [EP1442Y] (ab53013) at 1/1000 dilution
Lane 1 : Wild-type HeLa cell lysate
Lane 2 : ICAM1 CRISPR-Cas9 edited HeLa cell lysate
Lane 3 : A549 cell lysate
Lane 4 : Ramos cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 58 kDa
Observed band size: 90 kDa why is the actual band size different from the predicted?False colour image of Western blot: Anti-ICAM1 antibody [EP1442Y] staining at 1/1000 dilution, shown in green; Mouse anti-Alpha Tubulin [DM1A] (ab7291) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab53013 was shown to bind specifically to ICAM1. A band was observed at 90 kDa in wild-type HeLa cell lysates with no signal observed at this size in Icam1 knockout cell line ab261742 (knockout cell lysate ab256947). The band observed in the CRISPR-Cas9 edited lysate lane below 90 kDa (not observed by this antibody) is likely to represent a truncated form of ICAM1. This has not been investigated further and the functional properties of the gene product have not been determined. To generate this image, wild-type and Icam1 CRISPR-Cas9 edited HeLa cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween® 20 (TBS-T) 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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This data was developed using ab53013, the same antibody clone in a different buffer formulation.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human tonsil tissue sections labeling ICAM1 with purified ab53013 at 1/500 dilution (0.22 µg/mL). Heat mediated antigen retrieval using Bond™ Epitope Retrieval Solution 2 (pH 9.0). The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) was used as the secondary antibody. Negative control: PBS instead of the primary antibody. Hematoxylin was used as a counterstain. -
This data was developed using ab53013, the same antibody clone in a different buffer formulation.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human kidney tissue sections labeling ICAM1 with purified ab53013 at 1/500 dilution (0.22 µg/mL). Heat mediated antigen retrieval using Bond™ Epitope Retrieval Solution 2 (pH 9.0). The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) was used as the secondary antibody. Negative control: PBS instead of the primary antibody. Hematoxylin was used as a counterstain. -
All lanes : Anti-ICAM1 antibody [EP1442Y] - BSA and Azide free (ab271852) at 1/1000 dilution
Lane 1 : Untreated HUVEC (Human umbilical vein endothelial cell) whole cell lysate
Lane 2 : HUVEC (Human umbilical vein endothelial cell) treated with 50ng/ml TNF-a for 24 hours whole cell lysate
Lane 3 : Raji (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 (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 58 kDaThe molecular mass observed is consistent with what has been described in the literatures (PMID: 29777158, 30082828, 31244919).
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All lanes : Anti-ICAM1 antibody [EP1442Y] (ab53013) at 1/1000 dilution
Lane 1 : Wild-type HeLa cell lysate
Lane 2 : ICAM1 knockout HeLa cell lysate
Lane 3 : Raji cell lysate
Lane 4 : Ramos cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) at 1/10000 dilution
Predicted band size: 58 kDa
Observed band size: 90 kDa why is the actual band size different from the predicted?This data was developed using ab53013, the same antibody clone in a different buffer formulation.
Lanes 1-4: Merged signal (red and green). Green - ab53013 observed at 90 kDa. Red - loading control ab8245 observed at 36 kDa.
ab53013 Anti-ICAM1 antibody [EP1442Y] was shown to specifically react with ICAM1 in wild-type HeLa cells. 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. ab53013 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C 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 20000 dilution for 1 hour at room temperature before imaging.
实验方案
数据表及文件
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Datasheet download
Certificate of Compliance
文献 (14)
ab221777 被引用在 14 文献中.
- Shao X et al. Astragaloside IV alleviates atherosclerosis through targeting circ_0000231/miR-135a-5p/CLIC4 axis in AS cell model in vitro. Mol Cell Biochem 476:1783-1795 (2021). PubMed: 33439448
- Wei Y et al. MicroRNA-19b-3p suppresses gastric cancer development by negatively regulating neuropilin-1. Cancer Cell Int 20:193 (2020). PubMed: 32508529
- Shi H et al. Long non-coding RNA expression profile in minor salivary gland of primary Sjögren's syndrome. Arthritis Res Ther 18:109 (2016). IHC ; Human . PubMed: 27188286
- Broniarek I et al. The effect of chronic exposure to high palmitic acid concentrations on the aerobic metabolism of human endothelial EA.hy926 cells. Pflugers Arch 468:1541-54 (2016). WB ; Human . PubMed: 27417103
- Pannier D et al. High expression levels of egfl7 correlate with low endothelial cell activation in peritumoral vessels of human breast cancer. Oncol Lett 12:1422-1428 (2016). IHC-P ; Human . PubMed: 27446447