Thermo Fisher Scientific CD54 (ICAM-1) Monoclonal Antibody (1A29), PerCP-eFluor 710, eBioscience

Applications

Tested Dilution

Publications

Flow Cytometry (Flow)

0.125 µg/test

View 1 publication 1 publication

Product Specifications

Species Reactivity

Rat

Published species

Rat

Host/Isotype

Mouse / IgG1, kappa

Recommended Isotype Control

Mouse IgG1 kappa Isotype Control (P3.6.2.8.1), PerCP-eFluor™ 710, eBioscience™

Class

Monoclonal

Type

Antibody

Clone

1A29

Conjugate

PerCP-eFluor™ 710 PerCP-eFluor™ 710 PerCP-eFluor™ 710

Excitation/Emission Max

482/708 nm View spectra

Form

Liquid

Concentration

0.2 mg/mL

Purification

Affinity chromatography

Storage buffer

PBS, pH 7.2

Contains

0.09% sodium azide

Storage conditions

4° C, store in dark, DO NOT FREEZE!

Shipping conditions

Ambient (domestic); Wet ice (international)

RRID

AB_10854127

Product Specific Information

Description: The 1A29 monoclonal antibody reacts with rat CD54, which is also known as intercellular adhesion molecule 1 (ICAM-1). In the immune system, this type I immunoglobulin (Ig)-like transmembrane protein is expressed on lymphocytes, macrophages, dendritic cells, monocytes, granulocytes, and thymic stromal cells. CD54 is also present on endothelial cells and such non-immune cells as hepatocytes, cardiomyocytes, and vascular smooth muscle cells. In addition, reports demonstrate that CD54 expression can be induced in vitro by proinflammatory cytokines, including IFN-gamma, IL-1, CCL2, or TNF-alpha. CD54 plays important roles in transendothelial migration of leukocytes to sites of inflammation and in T cell interactions with antigen presenting cells. Studies indicate that CD54 mediates these functions by binding to lymphocyte function-associated molecule (LFA-1; also known as CD11a/CD18).

The 1A29 antibody has been reported to block interactions between CD54 and LFA-1.

Applications Reported: This 1A29 antibody has been reported for use in flow cytometric analysis.

Applications Tested: This 1A29 antibody has been tested by flow cytometric analysis of rat splenocytes. This can be used at less than or equal to 0.125 µg per test. A test is defined as the amount (µg) of antibody that will stain a cell sample in a final volume of 100 µL. Cell number should be determined empirically but can range from 10^5 to 10^8 cells/test. It is recommended that the antibody be carefully titrated for optimal performance in the assay of interest.

PerCP-eFluor® 710 emits at 710 nm and is excited with the blue laser (488 nm); it can be used in place of PerCP-Cyanine5.5. We recommend using a 710/50 bandpass filter, however, the 695/40 bandpass filter is an acceptable alternative. Please make sure that your instrument is capable of detecting this fluorochrome.

Fixation: Samples can be stored in IC Fixation Buffer (Product # 00-8222) (100 µL cell sample + 100 µL IC Fixation Buffer) or 1-step Fix/Lyse Solution (Product # 00-5333) for up to 3 days in the dark at 4°C with minimal impact on brightness and FRET efficiency/compensation. Some generalizations regarding fluorophore performance after fixation can be made, but clone specific performance should be determined empirically.

Excitation: 488 nm; Emission: 710 nm; Laser: Blue Laser.

Filtration: 0.2 µm post-manufacturing filtered.

Target Information

ICAM-1 (CD54) is an 85-110 kDa single-chain type 1 integral membrane glycoprotein with an extracellular domain of five immunoglobulin superfamily repeats, a transmembrane region and a cytoplasmic domain. ICAM-1 has 7 potential N-linked glycosylation sites and shares considerable amino acid sequence homology with ICAM-3 (CD50) and ICAM-2 (CD102). ICAM-1 binds to integrins of type CD11a/CD18 (leukocyte adhesion molecule, LFA-1), or CD11b/CD18 (Mac-1) and is exploited by Rhinovirus as a receptor. ICAM-1 is expressed by activated endothelial cells and detected on epithelial cells, fibroblasts, chondrocytes, B lymphocytes, T lymphocytes (low), monocytes, macrophages, dendritic cells and neutrophils, with lower levels that increase upon inflammation. ICAM-1 is also detected in some carcinoma and melanoma cells. Soluble ICAM-1 is detectable in the plasma and is elevated in patients with various inflammatory syndromes.

For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.