Recombinant Anti-ICAM1 antibody [EPR4776] (ab109361)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR4776] to ICAM1
- Suitable for: Flow Cyt (Intra), ICC/IF, WB, IP, IHC-P
- Reacts with: Human
Related conjugates and formulations
Overview
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Product name
Anti-ICAM1 antibody [EPR4776]
See all ICAM1 primary antibodies -
Description
Rabbit monoclonal [EPR4776] to ICAM1 -
Host species
Rabbit -
Tested applications
Suitable for: Flow Cyt (Intra), ICC/IF, WB, IP, IHC-Pmore details -
Species reactivity
Reacts with: Human -
Immunogen
Recombinant fragment within Human ICAM1 aa 300-500 (C terminal). The exact sequence is proprietary.
Database link: P05362 -
Positive control
- IHC-P: Human lung carcinoma and tonsil tissue sections; Flow Cyt (intra): Ramos cells; ICC/IF: Raji cells; IP: Raji whole cell lysate; WB: HUVEC treated with 10ng/ml for 18 hours and Ramos whole cell lysate.
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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 +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 0.05% BSA, 40% Glycerol (glycerin, glycerine), 59% PBS -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR4776 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Isotype control
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Positive Controls
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab109361 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 |
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Flow Cyt (Intra) |
1/70.
For unpurified use at 1/100 - 1/500. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
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ICC/IF |
1/100.
For unpurified use at 1/500. |
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WB |
1/1000 - 1/10000. Detects a band of approximately 89 kDa (predicted molecular weight: 58 kDa).
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IP |
1/10 - 1/100.
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IHC-P | (1) |
1/100 - 1/250. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
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Notes |
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Flow Cyt (Intra)
1/70. For unpurified use at 1/100 - 1/500. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
ICC/IF
1/100. For unpurified use at 1/500. |
WB
1/1000 - 1/10000. Detects a band of approximately 89 kDa (predicted molecular weight: 58 kDa). |
IP
1/10 - 1/100. |
IHC-P
1/100 - 1/250. Perform heat mediated antigen retrieval with citrate buffer pH 6 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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All lanes : Anti-ICAM1 antibody [EPR4776] (ab109361) 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 [EPR4776] 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, ab109361 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 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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Lane 1 : Anti-ICAM1 antibody [EPR4776] (ab109361) at 1/1000 dilution (Purified)
Lane 2 : Anti-ICAM1 antibody [EPR4776] (ab109361) at 1/1000 dilution ((Purified))
Lane 1 : HUVEC (Human umbilical vein endothelial cell) whole cell lysate
Lane 2 : HUVEC (Human umbilical vein endothelial cell) treated with 10ng/ml for 18 hours whole cell lysate
Lysates/proteins at 15 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution (Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated)
Predicted band size: 58 kDa -
Anti-ICAM1 antibody [EPR4776] (ab109361) at 1/1000 dilution (Purified) + Ramos (Human Burkitt's lymphoma B lymphocyte) whole cell lysate at 15 µg
Secondary
Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 58 kDa -
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ICAM1 antibody [EPR4776] (ab109361)Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human lung carcinoma tissue sections labeling ICAM1 with Purified ab109361 at 1/100 dilution (6.79 µg/mL). Heat mediated antigen retrieval using Bond™ Epitope Retrieval Solution 2 (pH 9.0). 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.
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Intracellular Flow Cytometry analysis of Ramos (Human Burkitt's lymphoma B lymphocyte) cells labeling ICAM1 with Purified ab109361 at 1/70 dilution (10 µg/mL) (Red). Cells were fixed with 4% Paraformaldehyde and permeabilised with 90% Methanol. A Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) secondary antibody was used at 1/2000. Isotype control - Rabbit monoclonal IgG (Black). Unlabeled control - Cell without incubation with primary antibody and secondary antibody (Blue).
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Purified ab109361 at 1/30 dilution (2µg) immunoprecipitating ICAM1 in Raji whole cell lysate.
Lane 1 (input): Raji (Human Burkitt's lymphoma B lymphocyte) whole cell lysate 10µg
Lane 2 (+): ab109361 + Raji whole cell lysate.
Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab109361 in Raji whole cell lysate.
VeriBlot for IP Detection Reagent (HRP) (ab131366) (1/1000 dilution) was used for Western blotting.
Blocking Buffer and concentration: 5% NFDM/TBST.
Diluting buffer and concentration: 5% NFDM/TBST.
Observed band size: 89 kDa -
Immunocytochemistry analysis of Raji (Human Burkitt's lymphoma B lymphocyte) cells labeling ICAM1 with Purified ab109361 at 1/100 dilution (6.79 µg/mL). Cells were fixed in 4% Paraformaldehyde and permeabilized with 0.1% tritonX-100. Cells were counterstained with Ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1/200 dilution (2.5 µg/mL). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) was used as the secondary antibody at 1/1000 dilution (2 µg/mL). DAPI (blue) was used as nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ICAM1 antibody [EPR4776] (ab109361)Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human tonsil tissue sections labeling ICAM1 with Purified ab109361 at 1/100 dilution (6.79 µg/mL). Heat mediated antigen retrieval using Bond™ Epitope Retrieval Solution 2 (pH 9.0). 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.
Protocols
Datasheets and documents
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SDS download
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Datasheet download
References (14)
ab109361 has been referenced in 14 publications.
- Li Q et al. Single-cell transcriptome profiling reveals vascular endothelial cell heterogeneity in human skin. Theranostics 11:6461-6476 (2021). PubMed: 33995668
- Yao MY et al. Long non-coding RNA MALAT1 exacerbates acute respiratory distress syndrome by upregulating ICAM-1 expression via microRNA-150-5p downregulation. Aging (Albany NY) 12:6570-6585 (2020). PubMed: 32315984
- Yang W et al. Potential role of lncRNA HULC/miR-128-3p/RAC1 axis in the inflammatory response during LPS-induced sepsis in HMEC-1 cells. Mol Med Rep 22:5095-5104 (2020). PubMed: 33174038
- Qiu D et al. Hyperglycemia Decreases Epithelial Cell Proliferation and Attenuates Neutrophil Activity by Reducing ICAM-1 and LFA-1 Expression Levels. Front Genet 11:616988 (2020). PubMed: 33414814
- Shishkova D et al. Calciprotein Particles Cause Endothelial Dysfunction under Flow. Int J Mol Sci 21:N/A (2020). PubMed: 33233811