Recombinant Anti-ICAM1 antibody [EPR4776] - BSA and Azide free (ab226059)
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 - BSA and Azide free
- Suitable for: Flow Cyt (Intra), WB, IP, IHC-P, ICC/IF
- Reacts with: Human
Related conjugates and formulations
Overview
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Product name
Anti-ICAM1 antibody [EPR4776] - BSA and Azide free
See all ICAM1 primary antibodies -
Description
Rabbit monoclonal [EPR4776] to ICAM1 - BSA and Azide free -
Host species
Rabbit -
Tested applications
Suitable for: Flow Cyt (Intra), WB, IP, IHC-P, ICC/IFmore 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
ab226059 is the carrier-free version of ab109361.
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 compatible with the Maxpar® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar® is a trademark of Fluidigm Canada Inc.
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.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C. Do Not Freeze. -
Storage buffer
pH: 7.2
Constituent: PBS -
Carrier free
Yes -
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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Compatible Secondaries
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Conjugation kits
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Isotype control
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Positive Controls
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab226059 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) |
Use at an assay dependent concentration.
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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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IP |
Use at an assay dependent concentration.
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IHC-P |
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
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ICC/IF |
Use at an assay dependent concentration.
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Notes |
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Flow Cyt (Intra)
Use at an assay dependent concentration. |
WB
Use at an assay dependent concentration. Detects a band of approximately 89 kDa (predicted molecular weight: 58 kDa). |
IP
Use at an assay dependent concentration. |
IHC-P
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol. |
ICC/IF
Use at an assay dependent concentration. |
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 knockout cell line ab261742 (knockout 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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This data was developed using ab109361, the same antibody clone in a different buffer formulation.
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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 kDaThis data was developed using ab109361, the same antibody clone in a different buffer formulation.
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This data was developed using ab109361, the same antibody clone in a different buffer formulation.
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. -
This data was developed using ab109361, the same antibody clone in a different buffer formulation. 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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This data was developed using ab109361, the same antibody clone in a different buffer formulation.
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 -
This data was developed using ab109361, the same antibody clone in a different buffer formulation.
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. -
This data was developed using ab109361, 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 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
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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Datasheet download
Certificate of Compliance
References (0)
ab226059 has not yet been referenced specifically in any publications.