ICAM-1/CD54 Polyclonal antibody

ICAM-1/CD54 Polyclonal Antibody for WB, IHC, IF/ICC, ELISA

Host / Isotype

Rabbit / IgG

Reactivity

human

Applications

WB, IHC, IF/ICC, ELISA

Conjugate

Unconjugated

Cat no : 30806-1-AP

Synonyms

CD54, ICAM-1, ICAM1, ICAM 1, BB2

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Validation Data Gallery

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  WB analysis using 30806-1-AP

  Various lysates were subjected to SDS PAGE followed by western blot with 30806-1-AP (ICAM-1 antibody) at dilution of 1:4000 incubated at room temperature for 1.5 hours.

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  IHC staining of human ovary cancer using 30806-1-AP

  Immunohistochemical analysis of paraffin-embedded human ovary cancer tissue slide using 30806-1-AP (ICAM-1/CD54 antibody) at dilution of 1:400 (under 10x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

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  IHC staining of human ovary cancer using 30806-1-AP

  Immunohistochemical analysis of paraffin-embedded human ovary cancer tissue slide using 30806-1-AP (ICAM-1/CD54 antibody) at dilution of 1:400 (under 20x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

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  IHC staining of human intrahepatic cholangiocarcinoma using 30806-1-AP

  Immunohistochemical analysis of paraffin-embedded human intrahepatic cholangiocarcinoma tissue slide using 30806-1-AP (ICAM-1/CD54 antibody) at dilution of 1:400 (under 20x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

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  IF Staining of Raji using 30806-1-AP

  Immunofluorescent analysis of (4% PFA) fixed Raji cells using ICAM-1 antibody (30806-1-AP) at dilution of 1:400 and CoraLite®488-Conjugated AffiniPure Goat Anti-Rabbit IgG(H+L) (SA00013-2).

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Tested Applications

• Positive WB detected in: Ramos cells, L02 cells, Raji cells
• Positive IHC detected in: human ovary cancer tissue, human intrahepatic cholangiocarcinoma tissue; Note: suggested antigen retrieval with TE buffer pH 9.0; (*) Alternatively, antigen retrieval may be performed with citrate buffer pH 6.0
• Positive IF/ICC detected in: Raji cells

Recommended dilution

• Western Blot (WB): WB : 1:1000-1:8000
• Immunohistochemistry (IHC): IHC : 1:200-1:800
• Immunofluorescence (IF)/ICC: IF/ICC : 1:200-1:800

It is recommended that this reagent should be titrated in each testing system to obtain optimal results.

Sample-dependent, Check data in validation data gallery.

Product Information

30806-1-AP targets ICAM-1/CD54 in WB, IHC, IF/ICC, ELISA applications and shows reactivity with human samples.

• Tested Reactivity: human
• Host / Isotype: Rabbit / IgG
• Class: Polyclonal
• Type: Antibody
• Immunogen: Recombinant protein
• Full Name: intercellular adhesion molecule 1
• Calculated Molecular Weight: 90 kDa
• Observed Molecular Weight: 85-95 kDa
• GenBank Accession Number: BC015969
• Gene Symbol: ICAM-1
• Gene ID (NCBI): 3383
• Conjugate: Unconjugated
• Form: Liquid
• Purification Method: Antigen affinity purification
• Storage Buffer: PBS with 0.02% sodium azide and 50% glycerol pH 7.3.
• Storage Conditions: Store at -20°C. Stable for one year after shipment. Aliquoting is unnecessary for -20^oC storage. 20ul sizes contain 0.1% BSA.

Background Information

Where is ICAM-1 expressed?

Intercellular Adhesion Molecule 1 (ICAM-1), also known as Cluster of Differentiation 54 (CD54) is a transmembrane glycoprotein constitutively expressed at low levels in endothelial cells, pericytes and on some lymphocytes and monocytes¹. It is located at the cytoplasmic membrane, with a large extracellular region of mainly hydrophobic amino acids joined to a small transmembrane region and a cytoplasmic tail. It has a molecular weight of 75 to 115 kDa depending on the level of glycosylation.

 What is the function of ICAM-1?

ICAM-1 is important in both innate and adaptive immune responses as an adhesion molecule. Although it is constitutively expressed, in the presence of pro-inflammatory cytokines such as TNFα the endothelial cells are activated and upregulate expression of ICAM-1². In blood vessels lined with endothelial cells, leukocytes that are rolling over the surface are able to bind to ICAM-1 and transmigrate through the endothelial barrier and into the tissue. The initial binding of the leukocytes to ICAM-1 causes a Ca2+ release that initiates endothelial cell contraction and weakening of the intercellular tight junctions^{3, 4}. This protein can be used as an indicator of endothelial activation and of vascular inflammation.

 What is the role of ICAM-1 in disease?

Beyond the role in the immune response, ICAM-1 has also been identified as the target of attachment for the human rhinovirus, the cause of the common cold. Binding of the virus to ICAM-1 causes the viral capsid to uncoat and leads to release of the genetic material⁵.

1.  Hubbard, A. K. & Rothlein, R. Intercellular adhesion molecule-1 (ICAM-1) expression and cell signaling cascades. Free Radic. Biol. Med. 28, 1379-86 (2000).

2. Long, E. O. ICAM-1: getting a grip on leukocyte adhesion. J. Immunol. 186, 5021-3 (2011).

3. Lawson, C. & Wolf, S. ICAM-1 signaling in endothelial cells. (2009).

4. Lyck, R. & Enzmann, G. The physiological roles of ICAM-1 and ICAM-2 in neutrophil migration into tissues. Curr. Opin. Hematol. 22, 53-59 (2015).

5. Xing, L., Casasnovas, J. M. & Cheng, R. H. Structural analysis of human rhinovirus complexed with ICAM-1 reveals the dynamics of receptor-mediated virus uncoating. J. Virol. 77, 6101-7 (2003).