QCM™ CHEMOTAXIS 96-WELL Cell Migration Assay Kit (3 µm Pore)

Key Applications	Detection Methods
ACT	Fluorescent

Key Applications

Detection Methods

ACT

Fluorescent

Description
Catalogue Number	ECM515
Brand Family	Chemicon®
Trade Name	QCM Chemicon
Description	QCM Chemotaxis Cell Migration Assay, 96-well (3 µm), fluorimetric
Overview	Also available: Cell Comb™ Scratch Assay! Get biochemical data from a scratch assay! Click Here Introduction Cell migration is a fundamental function of normal cellular processes, including embryonic development, angiogenesis, wound healing, immune response, and inflammation (1-2). One such process, leukocyte extravasation, is crucial for appropriate and effective immune response (3). Neutrophils normally exist in a resting state as they circulate though the body. However, upon interaction with small molecules known as chemoattractants, they rapidly respond with endothelial adhesion followed by emigration from the vasculature and chemotaxis to the site of inflammation (4-5). These chemoattractant receptors activate heterotrimeric GTP-binding proteins (G proteins) that initiate numerous elaborate signal transduction cascades, culminating in neutrophil migration and activation. Once at the site of inflammation, neutrophils respond with phagocytosis, superoxide generation, and the release of degradative enzymes. Microporous membrane inserts are widely used for cell migration and invasion assays. The most widely accepted of which is the Boyden Chamber assay. However, current methods of analysis are time-consuming and tedious, involving cotton swabbing of non-migrated cells on the topside of insert, manual staining and counting. Recently a fluorescence blocking membrane insert was introduced to address these issues; however, this approach requires labeling of the cells with Calcein-AM and extensive washing to remove free Calcein before cell migration. The effect of this treatment on cell behavior/migration remains questionable. The Chemicon QCM™ 96-well 3 μM Migration Assay does not require cell labeling, scraping, washing or counting. The 96-well insert and homogenous fluorescence detection format allows for large-scale screening and quantitative comparison of multiple samples (6). In the Chemicon QCM™ 3 μM 96-well Migration Assay, migratory cells on the bottom of the insert membrane are dissociated from the membrane when incubated with Cell Detachment Buffer. These cells are subsequently lysed and detected by the patented CyQuant GR dye (Molecular Probes) (7). This green-fluorescent dye exhibits strong fluorescence enhancement when bound to cellular nucleic acids (8). Most migration assays utilize an 8 μm pore size, as this is appropriate for most cell types, e.g. epithelial and fibroblast cells. The Chemicon QCM™ 3 μM 96-well Migration Assay utilizes a 3 μm pore size, which is appropriate for leukocyte migration. The system may be adapted to study different types of cell migration, including haptotaxis, random migration, chemokinesis, and chemotaxis. The Chemicon QCM™ 3 μM 96-well Migration Assay provides a quick and efficient system for quantitative determination of various factors on cell migration, including screening of pharmacological agents, evaluation of integrins or other adhesion receptors responsible for cell migration, or analysis of gene function in transfected cells. In addition, Chemicon also provides QCM™ 8μM 96-well Chemotaxis Cell Migration (ECM510) and Invasion Assays (ECM555) (8 μm pore), 24-well insert Cell Migration and Invasion Assay Systems, CytoMatrix™ Cell Adhesion strips coated with ECM proteins, or anti-integrin antibodies, and QuantiMatrix™ ECM protein ELISA kits. Application: The Chemicon QCM™ 96-well 3μM Migration Assay is ideal for studying chemotaxis cell migration. The quantitative nature of this assay is especially useful for large scale screening of pharmacological agents. The 3 μm pore size of this assay's Boyden chambers is appropriate for studying leukocyte migration. Each kit provides sufficient materials for the evaluation of 96 samples. The Chemicon QCM™ 3μM 96-well Migration Assay is intended for research use only; not for diagnostic applications.
Materials Required but Not Delivered	1. Precision pipettes: sufficient for aliquoting cells. 2. Harvesting buffer: EDTA or trypsin cell detachment buffer. Suggested formulations include a) 2 mM EDTA/PBS, b) 0.05% trypsin in Hanks Balanced Salt Solution (HBSS) containing 25 mM HEPES, or other cell detachment formulations as optimized by individual investigators. Note: Trypsin cell detachment buffer maybe required for difficult cell lines. Allow sufficient time for cell receptor recovery. 3. Tissue culture growth medium appropriate for subject cells, such as RPMI containing 10% FBS. 4. Chemoattractants (e.g. 10% FBS) or pharmacological agents for addition to culture medium, if screening is desired. 5. Quenching Medium: serum-free medium, such as RPMI, DMEM, or FBM (fibroblast basal media), containing 5% BSA. Note: Quenching Medium must contain divalent cations (Mg2+, Ca2+) sufficient for quenching EDTA in the harvesting buffer. 6. Sterile PBS or HBSS to wash cells. 7. Distilled water. 8. Low speed centrifuge and tubes for cell harvesting. 9. CO2 incubator appropriate for subject cells. 10. Hemocytometer or other means of counting cells. 11. Trypan blue or equivalent viability stain. 12. Fluorescence plate reader. 13. Sterile cell culture hood.

Description

Catalogue Number

ECM515

Brand Family

Chemicon®

Trade Name

QCM
Chemicon

Description

QCM Chemotaxis Cell Migration Assay, 96-well (3 µm), fluorimetric

Overview

Also available: Cell Comb™ Scratch Assay! Get biochemical data from a scratch assay! Click Here

Introduction
Cell migration is a fundamental function of normal cellular processes, including embryonic development, angiogenesis, wound healing, immune response, and inflammation (1-2). One such process, leukocyte extravasation, is crucial for appropriate and effective immune response (3). Neutrophils normally exist in a resting state as they circulate though the body. However, upon interaction with small molecules known as chemoattractants, they rapidly respond with endothelial adhesion followed by emigration from the vasculature and chemotaxis to the site of inflammation (4-5). These chemoattractant receptors activate heterotrimeric GTP-binding proteins (G proteins) that initiate numerous elaborate signal transduction cascades, culminating in neutrophil migration and activation. Once at the site of inflammation, neutrophils respond with phagocytosis, superoxide generation, and the release of degradative enzymes.

Microporous membrane inserts are widely used for cell migration and invasion assays. The most widely accepted of which is the Boyden Chamber assay. However, current methods of analysis are time-consuming and tedious, involving cotton swabbing of non-migrated cells on the topside of insert, manual staining and counting. Recently a fluorescence blocking membrane insert was introduced to address these issues; however, this approach requires labeling of the cells with Calcein-AM and extensive washing to remove free Calcein before cell migration. The effect of this treatment on cell behavior/migration remains questionable.

The Chemicon QCM™ 96-well 3 μM Migration Assay does not require cell labeling, scraping, washing or counting. The 96-well insert and homogenous fluorescence detection format allows for large-scale screening and quantitative comparison of multiple samples (6).

In the Chemicon QCM™ 3 μM 96-well Migration Assay, migratory cells on the bottom of the insert membrane are dissociated from the membrane when incubated with Cell Detachment Buffer. These cells are subsequently lysed and detected by the patented CyQuant GR dye (Molecular Probes) (7). This green-fluorescent dye exhibits strong fluorescence enhancement when bound to cellular nucleic acids (8).

Most migration assays utilize an 8 μm pore size, as this is appropriate for most cell types, e.g. epithelial and fibroblast cells. The Chemicon QCM™ 3 μM 96-well Migration Assay utilizes a 3 μm pore size, which is appropriate for leukocyte migration. The system may be adapted to study different types of cell migration, including haptotaxis, random migration, chemokinesis, and chemotaxis.

The Chemicon QCM™ 3 μM 96-well Migration Assay provides a quick and efficient system for quantitative determination of various factors on cell migration, including screening of pharmacological agents, evaluation of integrins or other adhesion receptors responsible for cell migration, or analysis of gene function in transfected cells.

In addition, Chemicon also provides QCM™ 8μM 96-well Chemotaxis Cell Migration (ECM510) and Invasion Assays (ECM555) (8 μm pore), 24-well insert Cell Migration and Invasion Assay Systems, CytoMatrix™ Cell Adhesion strips coated with ECM proteins, or anti-integrin antibodies, and QuantiMatrix™ ECM protein ELISA kits.

Application:

The Chemicon QCM™ 96-well 3μM Migration Assay is ideal for studying chemotaxis cell migration. The quantitative nature of this assay is especially useful for large scale screening of pharmacological agents. The 3 μm pore size of this assay's Boyden chambers is appropriate for studying leukocyte migration. Each kit provides sufficient materials for the evaluation of 96 samples.

The Chemicon QCM™ 3μM 96-well Migration Assay is intended for research use only; not for diagnostic applications.

Materials Required but Not Delivered

1. Precision pipettes: sufficient for aliquoting cells.

2. Harvesting buffer: EDTA or trypsin cell detachment buffer. Suggested formulations include a) 2 mM EDTA/PBS, b) 0.05% trypsin in Hanks Balanced Salt Solution (HBSS) containing 25 mM HEPES, or other cell detachment formulations as optimized by individual investigators.

Note: Trypsin cell detachment buffer maybe required for difficult cell lines. Allow sufficient time for cell receptor recovery.

3. Tissue culture growth medium appropriate for subject cells, such as RPMI containing 10% FBS.

4. Chemoattractants (e.g. 10% FBS) or pharmacological agents for addition to culture medium, if screening is desired.

5. Quenching Medium: serum-free medium, such as RPMI, DMEM, or FBM (fibroblast basal media), containing 5% BSA.

Note: Quenching Medium must contain divalent cations (Mg2+, Ca2+) sufficient for quenching EDTA in the harvesting buffer.

6. Sterile PBS or HBSS to wash cells.

7. Distilled water.

8. Low speed centrifuge and tubes for cell harvesting.

9. CO2 incubator appropriate for subject cells.

10. Hemocytometer or other means of counting cells.

11. Trypan blue or equivalent viability stain.

12. Fluorescence plate reader.

13. Sterile cell culture hood.

Product Information
Components	Sterile 96-well 3μM Cell Migration Plate Assembly: (Part No. 90199) One 96-well feeder tray, and one 96-well Cell Migration Chamber plate with 3 μm pore insert. Area = 0.3 cm², 50,000 cells per well 96-well Cell Culture Tray: (Part No. 90129) One 96-well feeder tray. Cell Detachment Solution: (Part No. 90131) One bottle - 16 mL. 4X Cell Lysis Buffer: (Part No. 90130) One bottle - 16 mL. CyQuant GR Dye®: (Part No. 90132) One vial - 75 μL
Detection method	Fluorescent

Product Information

Components

Sterile 96-well 3μM Cell Migration Plate Assembly: (Part No. 90199) One 96-well feeder tray, and one 96-well Cell Migration Chamber plate with 3 μm pore insert. Area = 0.3 cm², 50,000 cells per well
96-well Cell Culture Tray: (Part No. 90129) One 96-well feeder tray.
Cell Detachment Solution: (Part No. 90131) One bottle - 16 mL.
4X Cell Lysis Buffer: (Part No. 90130) One bottle - 16 mL.
CyQuant GR Dye®: (Part No. 90132) One vial - 75 μL

Detection method

Fluorescent

Applications
Application	The Chemicon QCM 3 uM 96-well Migration Assay provides a quick & efficient system for quantitative determination of various factors on cell migration, including screening of pharmacological agents or evaluation of integrins.
Key Applications	Activity Assay

Applications

Application

The Chemicon QCM 3 uM 96-well Migration Assay provides a quick & efficient system for quantitative determination of various factors on cell migration, including screening of pharmacological agents or evaluation of integrins.

Key Applications

Activity Assay

Biological Information
Species Reactivity	All

Biological Information

Species Reactivity

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.

Packaging Information
Material Size	1 plate
Material Package	96 wells

Packaging Information

Material Size

1 plate

Material Package

96 wells

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

实验服务

办公用品

QCM™ CHEMOTAXIS 96-WELL Cell Migration Assay Kit (3 µm Pore)

浏览了该商品的用户最终购买

浏览了该商品的用户还浏览了

重要规格表

我要咨询