Cell Adhesion Assay 细胞黏附_详细资料

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Introduction
Cell adhesion is a complex process involved in embryogenesis, migration/invasion, tissue remodeling,
and wound healing. To perform these processes, cells adhere to extracellular matrix components （via
adhesion receptors）, forming complexes with components of the cytoskeleton that ultimay affect cell
motility, differentiation, proliferation, and survival. The Cell Biolabs CytoSelect™ Cell Adhesion
Assay Kit provides a rapid, quantitative method for evaluating cell adhesion. The kit contains
sufficient reagents for the evaluation of 48 samples （40 Human Fibronectin-coated wells, 8 BSA-
coated wells）.

Assay Principle
The CytoSelect™ Cell Adhesion Assay Kit utilizes a Fibronectin-coated 48-well plate （see Adhesion
Plate Layout below）. First, cells are seeded onto the coated substrate, where the adherent cells are
captured. Next, unbound cells are washed away, and the adherent cells are fixed/stained. Finally, the
stain is extracted and quantified colorimetrically.

Related Products
1.   CBA-051: CytoSelect™ 48-Well Cell Adhesion Assay （Fibronectin-Coated, Fluorometric Format）
2.   CBA-052: CytoSelect™ 48-Well Cell Adhesion Assay （Collagen I-Coated, Colorimetric Format）
3.   CBA-060: CytoSelect™ 48-Well Cell Adhesion Assay （Collagen IV-Coated, Colorimetric Format）
4.   CBA-058: CytoSelect™ 48-Well Cell Adhesion Assay （Fibrinogen-Coated, Colorimetric Format）
5.   CBA-056: CytoSelect™ 48-Well Cell Adhesion Assay （Laminin-Coated, Colorimetric Format）
6.   CBA-070: CytoSelect™ 48-Well Cell Adhesion Assay （ECM Array, Colorimetric Format）

Kit Components
1.   Fibronectin Adhesion Plate （Part No. 105001）: One 48-well plate containing 40 Human
Fibronectin-coated wells and 8 BSA-coated wells （see layout below）
2.   Cell Stain Solution （Part No. 11002）: One Bottle – 10.0 mL
3.   Extraction Solution （Part No. 11003）: One Bottle – 10.0 mL

Adhesion Plate Layout
The layout below indicates the location of wells coated with Fibronectin and those coated with BSA.

2

1

2

3

4

5

6

7

8

A     Fibronectin       Fibronectin       Fibronectin        Fibronectin       Fibronectin        Fibronectin       Fibronectin        Fibronectin

B      Fibronectin        Fibronectin        Fibronectin        Fibronectin        Fibronectin        Fibronectin       Fibronectin        Fibronectin

C      Fibronectin        Fibronectin        Fibronectin        Fibronectin        Fibronectin        Fibronectin       Fibronectin        Fibronectin

D      Fibronectin        Fibronectin        Fibronectin        Fibronectin        Fibronectin        Fibronectin       Fibronectin        Fibronectin

E      Fibronectin        Fibronectin        Fibronectin        Fibronectin        Fibronectin        Fibronectin       Fibronectin        Fibronectin

F

BSA

BSA

BSA

BSA

BSA

BSA

BSA

BSA

Materials Not Supplied
1.   Cell culture medium
2.   Serum free medium, such as DMEM containing 0.5% BSA, 2 mM CaCl₂ and 2 mM MgCl₂
3.   Cell culture incubator （37ºC, 5% CO₂ atmosphere）
4.   1X PBS containing 2 mM CaCl2 and 2 mM MgCl2
5.   Light microscope
6.   96-well microtiter plate
7.   Microtiter plate reader

Storage
Store all kit components at 4ºC up to their expiration dates.

Assay Protocol
1.   Under sterile conditions, allow the Fibronectin Adhesion Plate to warm up at room temperature
for 10 minutes.
2.   Prepare a cell suspension containing 0.1-1.0 x 10⁶ cells/ml in serum free media. Agents that
inhibit or stimulate cell adhesion can be added directly to the cell suspension.
3.   Add 150 µL of the cell suspension to the inside of each well （BSA-coated wells are provided as
a negative control）.
4.   Incubate for 30-90 min in a cell culture incubator.
3

5.   Carefully discard or aspirate the media from each well （Note: Do not allow wells to dry）.
Gently wash each well 4-5 times with 250 µL PBS.
6.   Aspirate the PBS from each well and add 200 µL of Cell Stain Solution. Incubate for 10
minutes at room temperature.
7.   Discard or aspirate the Cell Stain Solution from the wells. Gently wash each well 4-5 times
with 500 µL deionized water.
8.   Discard the final wash and let the wells air dry.
9.   Add 200 µL of Extraction Solution per well, and then incubate 10 minutes on an orbital shaker.
10. Transfer 150 µL from each extracted sample to a 96-well microtiter plate and measure the OD
560nm in a plate reader.

Example of Results
The following figures demonstrate typical results with the CytoSelect™ 48-Well Cell Adhesion Assay
Kit. One should use the data below for reference only. This data should not be used to interpret actual
results.

2.5
MDA-231
HT-1080
HEK293
2

1.5

1

0.5

0
BSA           Col I         Col IV          FN             LN             VN

Figure 1. ECM-mediated Cell Adhesion. Serum starved cells were allowed to attach to ECM-
coated 48-well plate for 1 hr at 100,000 cells/well. Adherent cells were stained （left panel picture） and
quantified at OD 560nm after extraction （right panel figure）.

References
1.   Hynes, R. O. （1992） Cell 69:11-25.
2.   Schwartz, M. A., Schaller, M. D. and Ginsberg, M. H. （1995） Annu. Rev. Cell Dev. Biol. 11:549-
599.

4

Recent Product Citations
1.   Cervera, A.M. et al. （2008）. Cells silenced for SDHB expression display characteristic features of
the tumor phenotype. Cancer Res. 68:4058-4067.

Warranty
These products are warranted to perform as described in their labeling and in Cell Biolabs literature when used in
accordance with their instructions. THERE ARE NO WARRANTIES THAT EXTEND BEYOND THIS EXPRESSED
WARRANTY AND CELL BIOLABS DISCLAIMS ANY IMPLIED WARRANTY OF MERCHANTABILITY OR
WARRANTY OF FITNESS FOR PARTICULAR PURPOSE. CELL BIOLABS’ sole obligation and purchaser’s
exclusive remedy for breach of this warranty shall be, at the option of CELL BIOLABS, to repair or replace the products.
In no event shall CELL BIOLABS be liable for any proximate, incidental or consequential damages in connection with the
products.

2004-2008: Cell Biolabs, Inc. - All rights reserved. No part of these works may be reproduced in any form without
permissions in writing.

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