E-SELECTIN, ELISA KIT

Analytes Available	Species Reactivity	Key Applications	Detection Methods
sE-Selectin-1	H	ELISA	Chromogenic

Analytes Available

Species Reactivity

Key Applications

Detection Methods

sE-Selectin-1

ELISA

Chromogenic

Description
Catalogue Number	ECM330
Brand Family	Chemicon®
Trade Name	Chemicon
Description	Human sE-Selectin-1 ELISA
Overview	E-selectin (Endothelial Leukocyte Adhesion Molecule-1, ELAM-1) belongs to the selectin family of adhesion molecules [Hogg, 1992; Lasky, 1991]. Together with LECAM-1 (L-selectin) and GMP-140 (P-selectin), E-selectin mediates the initial interactions of leukocytes and platelets with endothelial cells [Shimizu et al., 1991; Vonandrian et al., 1992]. Molecular structure: The extracellular part of all selectins consists of an aminoterminal c-type lectin domain which specifically binds to carbohydrate ligands. This is followed by an EGF-like domain, and, in the case of E-selectin, by 6 short consensus repeats. The transmembrane portion of the molecule is followed by a short cytoplasmic tail. Selectins provide the first, loose contacts of polymorphonuclear cells with the endothelium in areas of inflammation. The binding partner for E-selectin contains sialyl LewisX oligosaccharide or lactosaminoglycans that contain either sialic acid or fucose [Foxall, et al., 1992; Parekh, 1991]. E-selectin is expressed on cytokine-activated endothelial cells, and promotes the adhesion of leukocytes to the endothelium. This initial binding event is a prerequisite for the activation of the immune cells via inflammatory mediators [Kyan-Aung et al., 1991; Lawrence & Springer, 1991]. In contrast to GMP-140, E-selectin is maximally expressed 2-4 hours after cell activation. During the following 24-48 hours E-selectin is shed from the cytoplasmic membrane into the circulation [Pigott et al., 1992]. The circulating form of this selectin attracts neutrophils and activates the b2-integrins in preparing the cells for migration. Soluble E-selectin levels could provide insights into several pathologies, including allergic reactions [Brasch & Sterry, 1992; Gundel et al., 1991; Mulligan et al., 1991], septic shock [Engelberts et al., 1992; Munro et al., 1991; Redl et al., 1991], vascular infection and inflammatory bowel disease [Koizumi et al., 1992]. Test Principle: An anti-E-selectin monoclonal antibody is adsorbed onto microwells. The pair of monoclonal antibodies used in this ELISA detect the soluble form of E-Selectin present in serum, plasma, urine, and other biological fluids. Soluble E-selectin present in a sample or standard then binds to antibodies adsorbed to the microwells. A second, HRP-conjugated monoclonal anti-E-selectin antibody is added and binds to E-selectin captured by the first antibody. Unbound enzyme-conjugated anti-E-selectin is removed with a wash step and HRP substrate solution is added to the wells. An amount of colored product is formed, proportional to the amount of soluble E-selectin present in the sample. The reaction is terminated by addition of acid and absorbance is measured at 450nm. A standard curve is prepared from six E-selectin standard dilutions and the E-selectin sample concentration is determined. Application: The E-selectin ELISA is an enzyme-linked immunosorbent assay for the quantitative detection of soluble Endothelial Leukocyte Adhesion Molecule-1 levels in cell culture supernatants, human serum, plasma, amniotic fluid, or other body fluids. The E-selectin ELISA is for research use only. Not for use in diagnostic or therapeutic procedures.
Alternate Names	CD62E
Materials Required but Not Delivered	- 5 mL and 10 mL graduated pipettes - 20 μL to 1000 μL adjustable single channel micropipettes with disposable tips - 50 μL to 300 μL adjustable multichannel micropipette with disposable tips - Multichannel micropipette reservoir - Beakers, flasks, cylinders necessary for preparation of reagents - Device for delivery of wash solution (multichannel wash bottle or automatic wash system) - Microwell strip reader capable of reading at 450 nm (620 nm as optional reference wavelength) - Glass-distilled or deionized water - Statistical calculator with program to perform linear regression analysis.

Description

Catalogue Number

ECM330

Brand Family

Chemicon®

Trade Name

Chemicon

Description

Human sE-Selectin-1 ELISA

Overview

E-selectin (Endothelial Leukocyte Adhesion Molecule-1, ELAM-1) belongs to the selectin family of adhesion molecules [Hogg, 1992; Lasky, 1991]. Together with LECAM-1 (L-selectin) and GMP-140 (P-selectin), E-selectin mediates the initial interactions of leukocytes and platelets with endothelial cells [Shimizu et al., 1991; Vonandrian et al., 1992].

Molecular structure: The extracellular part of all selectins consists of an aminoterminal c-type lectin domain which specifically binds to carbohydrate ligands. This is followed by an EGF-like domain, and, in the case of E-selectin, by 6 short consensus repeats. The transmembrane portion of the molecule is followed by a short cytoplasmic tail.

Selectins provide the first, loose contacts of polymorphonuclear cells with the endothelium in areas of inflammation. The binding partner for E-selectin contains sialyl LewisX oligosaccharide or lactosaminoglycans that contain either sialic acid or fucose [Foxall, et al., 1992; Parekh, 1991]. E-selectin is expressed on cytokine-activated endothelial cells, and promotes the adhesion of leukocytes to the endothelium. This initial binding event is a prerequisite for the activation of the immune cells via inflammatory mediators [Kyan-Aung et al., 1991; Lawrence & Springer, 1991]. In contrast to GMP-140, E-selectin is maximally expressed 2-4 hours after cell activation. During the following 24-48 hours E-selectin is shed from the cytoplasmic membrane into the circulation [Pigott et al., 1992]. The circulating form of this selectin attracts neutrophils and activates the b2-integrins in preparing the cells for migration.

Soluble E-selectin levels could provide insights into several pathologies, including allergic reactions [Brasch & Sterry, 1992; Gundel et al., 1991; Mulligan et al., 1991], septic shock [Engelberts et al., 1992; Munro et al., 1991; Redl et al., 1991], vascular infection and inflammatory bowel disease [Koizumi et al., 1992].

Test Principle:

An anti-E-selectin monoclonal antibody is adsorbed onto microwells.

The pair of monoclonal antibodies used in this ELISA detect the soluble form of E-Selectin present in serum, plasma, urine, and other biological fluids.

Soluble E-selectin present in a sample or standard then binds to antibodies adsorbed to the microwells. A second, HRP-conjugated monoclonal anti-E-selectin antibody is added and binds to E-selectin captured by the first antibody.

Unbound enzyme-conjugated anti-E-selectin is removed with a wash step and HRP substrate solution is added to the wells.

An amount of colored product is formed, proportional to the amount of soluble E-selectin present in the sample. The reaction is terminated by addition of acid and absorbance is measured at 450nm. A standard curve is prepared from six E-selectin standard dilutions and the E-selectin sample concentration is determined.

Application:

The E-selectin ELISA is an enzyme-linked immunosorbent assay for the quantitative detection of soluble Endothelial Leukocyte Adhesion Molecule-1 levels in cell culture supernatants, human serum, plasma, amniotic fluid, or other body fluids. The E-selectin ELISA is for research use only. Not for use in diagnostic or therapeutic procedures.

Alternate Names

CD62E

Materials Required but Not Delivered

- 5 mL and 10 mL graduated pipettes

- 20 μL to 1000 μL adjustable single channel micropipettes with

disposable tips

- 50 μL to 300 μL adjustable multichannel micropipette with disposable tips

- Multichannel micropipette reservoir

- Beakers, flasks, cylinders necessary for preparation of reagents

- Device for delivery of wash solution (multichannel wash bottle or

automatic wash system)

- Microwell strip reader capable of reading at 450 nm (620 nm as

optional reference wavelength)

- Glass-distilled or deionized water

- Statistical calculator with program to perform linear regression

analysis.

Product Information
Components	· 1 aluminum pouches with a Microwell Plate coated with Monoclonal Antibody (murine) to human E-Selectin · 1 vial (200 μL) HRP-Conjugated Anti-E-Selectin Monoclonal (murine) Antibody · 2 vials (100 ng/mL each, reconstituted) soluble E-selectin Standard, lyophilized {dissolve in 390 microliters for lot 0606032906}. · 1 vial lyophilized control serum · 1 bottle (50 mL) Wash Buffer Concentrate 20X (PBS with 1% Tween 20)* · 1 vial (5 mL) Assay Buffer Concentrate 20X (PBS with 1% Tween 20 and 10% BSA)* · 1 bottle (12 mL) Sample Diluent (buffered protein matrix)* · 1 vial (7 mL) Substrate Solution I (tetramethyl-benzidine) · 1 vial (7 mL) Substrate Solution II (0.02 % buffered hydrogen peroxide) · 1 vial (12 mL) Stop Solution (1M Phosphoric Acid) · 1 Microwell Strip Holder · 2 Adhesive Plate Covers * reagents containing 0.01% thimerosal as preservative
Detection method	Chromogenic

Product Information

Components

· 1 aluminum pouches with a Microwell Plate coated with Monoclonal Antibody (murine) to human E-Selectin
· 1 vial (200 μL) HRP-Conjugated Anti-E-Selectin Monoclonal (murine) Antibody
· 2 vials (100 ng/mL each, reconstituted) soluble E-selectin Standard, lyophilized {dissolve in 390 microliters for lot 0606032906}.
· 1 vial lyophilized control serum
· 1 bottle (50 mL) Wash Buffer Concentrate 20X (PBS with 1%
Tween 20)*
· 1 vial (5 mL) Assay Buffer Concentrate 20X (PBS with 1% Tween 20 and 10% BSA)*
· 1 bottle (12 mL) Sample Diluent (buffered protein matrix)*
· 1 vial (7 mL) Substrate Solution I (tetramethyl-benzidine)
· 1 vial (7 mL) Substrate Solution II (0.02 % buffered hydrogen peroxide)
· 1 vial (12 mL) Stop Solution (1M Phosphoric Acid)
· 1 Microwell Strip Holder
· 2 Adhesive Plate Covers
* reagents containing 0.01% thimerosal as preservative

Detection method

Chromogenic

Applications
Application	The E-selectin ELISA quantitates soluble Endothelial Leukocyte Adhesion Molecule-1 levels in cell culture supernatants, human serum, plasma, amniotic fluid, or other body fluids.
Key Applications	ELISA

Applications

Application

The E-selectin ELISA quantitates soluble Endothelial Leukocyte Adhesion Molecule-1 levels in cell culture supernatants, human serum, plasma, amniotic fluid, or other body fluids.

Key Applications

ELISA

Biological Information
Species Reactivity	Human
Analytes Available	sE-Selectin-1
Entrez Gene Number	NM_000450.1
Entrez Gene Summary	The endothelial leukocyte adhesion molecule-1 is expressed by cytokine-stimulated endothelial cells. It is thought to be responsible for the accumulation of blood leukocytes at sites of inflammation by mediating the adhesion of cells to the vascular lining. It exhibits structural features such as the presence of lectin- and EGF-like domains followed by short consensus repeat (SCR) domains that contain 6 conserved cysteine residues. These proteins are part of the selectin family of cell adhesion molecules. This gene is present in single copy in the human genome and contains 14 exons spanning about 13 kb of DNA. Adhesion molecules participate in the interaction between leukocytes and the endothelium and appear to be involved in the pathogenesis of atherosclerosis.
Gene Symbol	SELE LECAM2 ESEL CD62E ELAM1 ELAM ELAM-1
UniProt Number	P16581
UniProt Summary	FUNCTION: SwissProt: P16581 # Expressed on cytokine induced endothelial cells and mediates their binding to leukocytes. The ligand recognized by ELAM-1 is sialyl-lewis X (alpha(1->3)fucosylated derivatives of polylactosamine that are found at the nonreducing termini of glycolipids). SIZE: 610 amino acids; 66655 Da SUBCELLULAR LOCATION: Membrane; Single-pass type I membrane protein. SIMILARITY: SwissProt: P16581 ## Belongs to the selectin/LECAM family. & Contains 1 C-type lectin domain. & Contains 1 EGF-like domain. & Contains 6 Sushi (CCP/SCR) domains.

Biological Information

Species Reactivity

Human

Analytes Available

sE-Selectin-1

Entrez Gene Number

NM_000450.1

Entrez Gene Summary

The endothelial leukocyte adhesion molecule-1 is expressed by cytokine-stimulated endothelial cells. It is thought to be responsible for the accumulation of blood leukocytes at sites of inflammation by mediating the adhesion of cells to the vascular lining. It exhibits structural features such as the presence of lectin- and EGF-like domains followed by short consensus repeat (SCR) domains that contain 6 conserved cysteine residues. These proteins are part of the selectin family of cell adhesion molecules. This gene is present in single copy in the human genome and contains 14 exons spanning about 13 kb of DNA. Adhesion molecules participate in the interaction between leukocytes and the endothelium and appear to be involved in the pathogenesis of atherosclerosis.

Gene Symbol

SELE
LECAM2
ESEL
CD62E
ELAM1
ELAM
ELAM-1

UniProt Number

P16581

UniProt Summary

FUNCTION: SwissProt: P16581 # Expressed on cytokine induced endothelial cells and mediates their binding to leukocytes. The ligand recognized by ELAM-1 is sialyl-lewis X (alpha(1->3)fucosylated derivatives of polylactosamine that are found at the nonreducing termini of glycolipids).
SIZE: 610 amino acids; 66655 Da
SUBCELLULAR LOCATION: Membrane; Single-pass type I membrane protein.
SIMILARITY: SwissProt: P16581 ## Belongs to the selectin/LECAM family. & Contains 1 C-type lectin domain. & Contains 1 EGF-like domain. & Contains 6 Sushi (CCP/SCR) domains.

Product Usage Statements
Usage Statement	Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.

Product Usage Statements

Usage Statement

Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.

Storage and Shipping Information
Storage Conditions	Storage of Kit Components Store kit reagents between 2° and 8°C. Store the lyophilized control at -20°C. Immediately after use remaining reagents should be returned to cold storage (2° to 8°C) with the exception of reconstituted control (-20°C). Expiration date of the kit and reagents is stated on the kit box. Precautions: · Reagents are intended for research use only and are not for use in diagnostic or therapeutic procedures. · Do not mix or substitute reagents with those from other lots or other sources. · Do not use kit reagents beyond expiration date on label. · Do not expose kit reagents to strong light during storage or incubation. · Rubber or disposable latex gloves should be worn while handling kit reagents or specimens. · Some reagents contain thimerosal as preservative, which is highly toxic by inhalation, ingestion, or contact with skin. Thimerosal is a possible mutagen and should be handled accordingly. · Avoid contact of substrate solutions with oxidizing agents and metal. · In order to avoid microbial contamination or cross-contamination of reagents or specimens which may invalidate the test use disposable pipette tips and/or pipettes. · Use clean, dedicated reagent trays for dispensing the conjugate and substrate reagents. · Exposure to acids will inactivate the conjugate. · Glass-distilled water or deionized water must be used for reagent preparation. · Substrate solutions must be at room temperature prior to use.

Storage and Shipping Information

Storage Conditions

Storage of Kit Components
Store kit reagents between 2° and 8°C. Store the lyophilized control at -20°C. Immediately after use remaining reagents should be returned to cold storage (2° to 8°C) with the exception of reconstituted control (-20°C). Expiration date of the kit and reagents is stated on the kit box.

Precautions:

· Reagents are intended for research use only and are not for use in diagnostic or therapeutic procedures.

· Do not mix or substitute reagents with those from other lots or other sources.

· Do not use kit reagents beyond expiration date on label.

· Do not expose kit reagents to strong light during storage or

incubation.

· Rubber or disposable latex gloves should be worn while handling kit reagents or specimens.

· Some reagents contain thimerosal as preservative, which is highly toxic by inhalation, ingestion, or contact with skin. Thimerosal is a possible mutagen and should be handled accordingly.

· Avoid contact of substrate solutions with oxidizing agents and metal.

· In order to avoid microbial contamination or cross-contamination of reagents or specimens which may invalidate the test use disposable pipette tips and/or pipettes.

· Use clean, dedicated reagent trays for dispensing the conjugate and substrate reagents.

· Exposure to acids will inactivate the conjugate.

· Glass-distilled water or deionized water must be used for reagent

preparation.

· Substrate solutions must be at room temperature prior to use.

Packaging Information
Material Size	1 kit

Packaging Information

Material Size

1 kit

实验耗材

分子生物学试剂

蛋白与免疫学

生化试剂

细胞及检测试剂

色谱及层析

仪器设备

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