Recombinant Anti-ICAM1 antibody [EP1442Y] (ab53013)
![Western blot - Anti-ICAM1 antibody [EP1442Y] (ab53013)](https://www.abcam.com/ps/products/53/ab53013/Images/ab53013-479672-anti-icam1-antibody-ep1442y-western-blot-wildtype-hela-icam1-knockout-hela-a549-ramos-ab.jpg "Western blot - Anti-ICAM1 antibody [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
Overview
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Product name
Anti-ICAM1 antibody [EP1442Y]
See all ICAM1 primary antibodies -
Description
Rabbit monoclonal [EP1442Y] to ICAM1 -
Host species
Rabbit -
Tested applications
Suitable for: WB, IHC-Pmore details -
Species reactivity
Reacts with: Human -
Immunogen
Synthetic peptide within Human ICAM1 aa 1-100 (N terminal). The exact sequence is proprietary.
(Peptide available asab218845) -
Positive control
- WB: Huvec, Ramos and Raji whole cell lysate; IHC: Human kidney and tonsil tissue.
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General notes
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.
We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.
Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at -20°C. Stable for 12 months at -20°C. -
Storage buffer
pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA -
Concentration information loading... -
Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EP1442Y -
Isotype
IgG -
Research areas
Associated products
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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
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KO cell pellets
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab53013 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
| Application | Abreviews | Notes |
|---|---|---|
| WB |
1/1000. Detects a band of approximately 89 kDa (predicted molecular weight: 58 kDa).
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| IHC-P |
1/500. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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| Notes |
|---|
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WB
1/1000. Detects a band of approximately 89 kDa (predicted molecular weight: 58 kDa). |
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IHC-P
1/500. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
Target
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Function
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. -
Sequence similarities
Belongs to the immunoglobulin superfamily. ICAM family.
Contains 5 Ig-like C2-type (immunoglobulin-like) domains. -
Post-translational
modificationsMonoubiquitinated, which is promoted by MARCH9 and leads to endocytosis. -
Cellular localization
Membrane. - Information by UniProt
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Database links
- Entrez Gene: 3383 Human
- Omim: 147840 Human
- SwissProt: P05362 Human
- Unigene: 643447 Human
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Alternative names
- Antigen identified by monoclonal antibody BB2 antibody
- BB 2 antibody
- BB2 antibody
see all
Images
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Western blot - Anti-ICAM1 antibody [EP1442Y] (ab53013)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.
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![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ICAM1 antibody [EP1442Y] (ab53013)](https://www.abcam.com/ps/products/53/ab53013/Images/ab53013-10-anti-icam1-antibody-ep1442y-immunohistochemistry-tonsil-human.jpg)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ICAM1 antibody [EP1442Y] (ab53013)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) - Anti-ICAM1 antibody [EP1442Y] (ab53013)](https://www.abcam.com/ps/products/53/ab53013/Images/ab53013-9-anti-icam1-antibody-ep1442y-immunohistochemistry-kidney-human.jpg)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ICAM1 antibody [EP1442Y] (ab53013)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. -
![Western blot - Anti-ICAM1 antibody [EP1442Y] (ab53013)](https://www.abcam.com/ps/products/53/ab53013/Images/ab53013-458210-anti-icam1-antibody-ep1442y-westernblot-human.jpg)
Western blot - Anti-ICAM1 antibody [EP1442Y] (ab53013)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-α for 24 hours whole cell lysate
Lane 3 : Raji (Human Burkitt's lymphoma B lymphocyte) whole cell lysate
Secondary
Lanes 1 & 3 : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/20000 dilution
Lane 2 : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution (Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated)
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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![Western blot - Anti-ICAM1 antibody [EP1442Y] (ab53013)](https://www.abcam.com/ps/products/53/ab53013/Images/ab53013-410190-anti-icam1-antibody-ep1442y-western-blot-human-knockout-lysate.jpg)
Western blot - Anti-ICAM1 antibody [EP1442Y] (ab53013)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.
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![Anti-ICAM1 antibody [EP1442Y] (ab53013)](https://www.abcam.com/ps/products/53/ab53013/Images/ab53013-7-benefits-of-recombinant-antibodies.png)
Anti-ICAM1 antibody [EP1442Y] (ab53013)
Protocols
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.
Datasheets and documents
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SDS download
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Datasheet download
References (46)
ab53013 has been referenced in 46 publications.
- Pars S et al. Generation of Functional Vascular Endothelial Cells and Pericytes from Keratinocyte Derived Human Induced Pluripotent Stem Cells. Cells 10:N/A (2021). PubMed: 33466396
- Wang K et al. Vertebral-specific activation of the CX3CL1/ICAM-1 signaling network mediates non-small-cell lung cancer spinal metastasis by engaging tumor cell-vertebral bone marrow endothelial cell interactions. Theranostics 11:4770-4789 (2021). PubMed: 33754027
- Wei Q et al. TNFa secreted by glioma associated macrophages promotes endothelial activation and resistance against anti-angiogenic therapy. Acta Neuropathol Commun 9:67 (2021). PubMed: 33853689
- Williams H et al. Aneurysm severity is suppressed by deletion of CCN4. J Cell Commun Signal 15:421-432 (2021). PubMed: 34080128
- Li X et al. Circ0120816 acts as an oncogene of esophageal squamous cell carcinoma by inhibiting miR-1305 and releasing TXNRD1. Cancer Cell Int 20:526 (2020). PubMed: 33292234