Anti-ICAM1 抗体 [EP1442Y] (ab53013)
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
- Suitable for: WB, IHC-P
- Reacts with: Human
Related conjugates and formulations
製品の概要
-
製品名
Anti-ICAM1 antibody [EP1442Y]
ICAM1 一次抗体 製品一覧 -
製品の詳細
Rabbit monoclonal [EP1442Y] to ICAM1 -
由来種
Rabbit -
アプリケーション
適用あり: WB, IHC-Pmore details -
種交差性
交差種: Human -
免疫原
Synthetic peptide within Human ICAM1 aa 1-100 (N terminal). The exact sequence is proprietary.
(Peptide available asab218845) -
ポジティブ・コントロール
- WB: Huvec, Ramos and Raji whole cell lysate; IHC: Human kidney and tonsil tissue.
-
特記事項
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.
Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.
製品の特性
-
製品の状態
Liquid -
保存方法
Shipped at 4°C. Store at -20°C. Stable for 12 months at -20°C. -
バッファー
pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA -
Concentration information loading...
-
精製度
Protein A purified -
ポリ/モノ
モノクローナル -
クローン名
EP1442Y -
アイソタイプ
IgG -
研究分野
関連製品
-
Alternative Versions
-
Compatible Secondaries
-
Isotype control
-
KO cell lines
-
KO cell lysates
アプリケーション
The Abpromise guarantee
Abpromise保証は、 次のテスト済みアプリケーションにおけるab53013の使用に適用されます
アプリケーションノートには、推奨の開始希釈率がありますが、適切な希釈率につきましてはご検討ください。
アプリケーション | Abreviews | 特記事項 |
---|---|---|
WB |
1/1000. Detects a band of approximately 89 kDa (predicted molecular weight: 58 kDa).
|
|
IHC-P |
1/500. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
|
特記事項 |
---|
WB
1/1000. Detects a band of approximately 89 kDa (predicted molecular weight: 58 kDa). |
IHC-P
1/500. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
ターゲット情報
-
機能
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. -
配列類似性
Belongs to the immunoglobulin superfamily. ICAM family.
Contains 5 Ig-like C2-type (immunoglobulin-like) domains. -
翻訳後修飾
Monoubiquitinated, which is promoted by MARCH9 and leads to endocytosis. -
細胞内局在
Membrane. - Information by UniProt
-
参照データベース
- Entrez Gene: 3383 Human
- Omim: 147840 Human
- SwissProt: P05362 Human
- Unigene: 643447 Human
-
別名
- Antigen identified by monoclonal antibody BB2 antibody
- BB 2 antibody
- BB2 antibody
see all
画像
-
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 CRISPR-Cas9 edited cell line ab261742 (CRISPR-Cas9 edited 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.
-
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.
-
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.
-
Lanes 1 & 3 : Anti-ICAM1 antibody [EP1442Y] (ab53013) at 1/1000 dilution
Lane 2 : Anti-ICAM1 antibody [EP1442Y] (ab53013) at 1/1000 dilution (Purified)
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).
-
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?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.
プロトコール
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.
データシートおよび資料
-
SDS download
-
Datasheet download
参考文献 (56)
ab53013 は 56 報の論文で使用されています。
- Williams D et al. Stable flow-induced expression of KLK10 inhibits endothelial inflammation and atherosclerosis. Elife 11:N/A (2022). PubMed: 35014606
- Jhang JF et al. Improved Urothelial Cell Proliferation, Cytoskeleton and Barrier Function Protein Expression in the Patients With Interstitial Cystitis/Bladder Pain Syndrome After Intravesical Platelet-Rich Plasma Injection. Int Neurourol J 26:S57-67 (2022). PubMed: 35073671
- Yang Y et al. A functional SNP rs895819 on pre-miR-27a is associated with bipolar disorder by targeting NCAM1. Commun Biol 5:309 (2022). PubMed: 35379867
- Versele R et al. TNF-α and IL-1β Modulate Blood-Brain Barrier Permeability and Decrease Amyloid-β Peptide Efflux in a Human Blood-Brain Barrier Model. Int J Mol Sci 23:N/A (2022). PubMed: 36142143
- Huang J et al. Micro RNA miR-1303 Promotion of Proliferation, Migration and Invasion of Human Liver Cancer Cells Is Enhanced by Low Talin 1 (TLN1) Expression. Anticancer Res 42:4715-4725 (2022). PubMed: 36192016