Recombinant Anti-ICAM1 antibody [EPR22161-284] (ab222736)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR22161-284] to ICAM1
- Suitable for: WB, IHC-P, Flow Cyt, ICC/IF, IP
- Reacts with: Mouse
Related conjugates and formulations
Overview
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Product name
Anti-ICAM1 antibody [EPR22161-284]
See all ICAM1 primary antibodies -
Description
Rabbit monoclonal [EPR22161-284] to ICAM1 -
Host species
Rabbit -
Tested applications
Suitable for: WB, IHC-P, Flow Cyt, ICC/IF, IPmore details -
Species reactivity
Reacts with: Mouse -
Immunogen
Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: Mouse kidney, spleen and liver tissue lysate; A20 whole cell lysate. IHC-P: Mouse lung and spleen tissue. ICC/IF: A20 cells. Flow: Mouse primary splenocytes. IP: Mouse spleen 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.
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: 7.2
Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR22161-284 -
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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Related Products
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab222736 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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WB |
1/1000. Detects a band of approximately 110 kDa (predicted molecular weight: 57 kDa).
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IHC-P |
1/1000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins. |
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Flow Cyt |
1/500.
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ICC/IF |
1/50.
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IP |
1/30.
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Notes |
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WB
1/1000. Detects a band of approximately 110 kDa (predicted molecular weight: 57 kDa). |
IHC-P
1/1000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins. |
Flow Cyt
1/500. |
ICC/IF
1/50. |
IP
1/30. |
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: 15894 Mouse
- SwissProt: P13597 Mouse
- Unigene: 435508 Mouse
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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 [EPR22161-284] (ab222736) at 1/1000 dilution
Lane 1 : Mouse kidney tissue lysate
Lane 2 : Mouse spleen tissue lysate
Lane 3 : Mouse liver tissue lysate
Lane 4 : A20 (mouse reticulum sarcoma b lymphocyte) whole cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Predicted band size: 57 kDa
Observed band size: 110 kDa why is the actual band size different from the predicted?Blocking/Diluting buffer and concentration: 5% NFDM/TBST.
Exposure time: 48 seconds.
The 110-KDa apparent molecular mass is due to glycosylation (PMID: 1680698).
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Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized A20 (mouse reticulum sarcoma cell line) cells labeling ICAM with ab222736 at 1/50 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing membranous staining in A20 cells with distinct staining of the target protein being transported from Golgi to cell surfaces (PMID: 17486117) is observed. Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) (ab195889) was used as a countertsain (Red) at 1/200 dilution. The nuclear counterstain is DAPI (Blue).
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Alexa Fluor®488 Goat anti-Rabbit secondary (ab150077) at 1/1000 dilution.
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Immunohistochemical analysis of paraffin-embedded mouse spleen tissue labeling ICAM1 using ab222736 at 1/1000 dilution, followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Cytoplasmic staining on the marginal zone of mouse spleen (PMID: 12130787; PMID: 17947659) is observed. Counterstained with Hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) ready to use.
Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.
Immunostaining was performed on a Leica Biosystems BOND® RX instrument. The section was incubated with ab222736 for 30 mins at room temperature.
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ICAM1 was immunoprecipitated from 0.35 mg Mouse spleen lysate with ab222736 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab222736 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366) was used for detection at 1/5000 dilution.
Lane 1: Mouse spleen tissue lysate 10 μg (input).
Lane 2: ab222736 IP in mouse spleen tissue lysate.
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab222736 in Mouse spleen lysate.Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 3 minutes.110-kDa molecular mass is due to glycosylation (PMID: 1680698).
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Flow cytometric analysis of Mouse primary splenocytes labeling ICAM1 with ab222736 at 1/500 dilution (Red) compared with a Rabbit monoclonal IgG (ab172730) isotype control (Black) and an unlabeled control (Cell without incubation with primary antibody and secondary antibody, Blue). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) at 1/200 dilution was used as the secondary antibody. Gated on viable cells.
ICAM1 negative population (left half of red peak) was also observed in mouse splenocytes which is consistent with IHC staining pattern.
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Immunohistochemical analysis of paraffin-embedded mouse lung tissue labeling ICAM1 using ab222736 at 1/1000 dilution, followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Membranous and cytoplasmic staining on mouse lung (PMID: 14568945; PMID: 9802985) is observed. Counterstained with DAPI.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) ready to use.
Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.
Immunostaining was performed on a Leica Biosystems BOND® RX instrument. The section was incubated with ab222736 for 30 mins at room temperature.
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
Certificate of Compliance
References (10)
ab222736 has been referenced in 10 publications.
- Qiao T et al. Combined pembrolizumab and bevacizumab therapy effectively inhibits non-small-cell lung cancer growth and prevents postoperative recurrence and metastasis in humanized mouse model. Cancer Immunol Immunother 72:1169-1181 (2023). PubMed: 36357599
- Wu D et al. Targeting uridine-cytidine kinase 2 induced cell cycle arrest through dual mechanism and could improve the immune response of hepatocellular carcinoma. Cell Mol Biol Lett 27:105 (2022). PubMed: 36447138
- Wang F et al. Single-Cell Sequencing Reveals the Regulatory Role of Maresin1 on Neutrophils during Septic Lung Injury. Cells 11:N/A (2022). PubMed: 36496993
- Liu Y et al. Deciphering the combination mechanisms of Gualou-Xiebai herb pair against atherosclerosis by network pharmacology and HPLC-Q-TOF-MS technology. Front Pharmacol 13:941400 (2022). PubMed: 36120369
- Bae SH et al. Neutrophils infiltrate into the spiral ligament but not the stria vascularis in the cochlea during lipopolysaccharide-induced inflammation. Theranostics 11:2522-2533 (2021). PubMed: 33456557
- Shi FJ et al. Is Iba-1 protein expression a sensitive marker for microglia activation in experimental diabetic retinopathy? Int J Ophthalmol 14:200-208 (2021). PubMed: 33614447
- Xu XH et al. Interleukin-6 released by oral lichen planus myofibroblasts promotes angiogenesis. Exp Ther Med 21:291 (2021). PubMed: 33717234
- Lv P et al. SM22a Loss Contributes to Apoptosis of Vascular Smooth Muscle Cells via Macrophage-Derived circRasGEF1B. Oxid Med Cell Longev 2021:5564884 (2021). PubMed: 33859778
- Tong Y et al. The β3/5 Integrin-MMP9 Axis Regulates Pulmonary Inflammatory Response and Endothelial Leakage in Acute Lung Injury. J Inflamm Res 14:5079-5094 (2021). PubMed: 34675589
- Wu W et al. Lnc-SNHG16/miR-128 axis modulates malignant phenotype through WNT/ß-catenin pathway in cervical cancer cells. J Cancer 11:2201-2212 (2020). PubMed: 32127947