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48-602MAG
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The Alpha/Beta Integrin-Mediated Cell Adhesion Array Kit can be used for assessing the presence or absence of specific integrins on the cell surface.
More>>The Alpha/Beta Integrin-Mediated Cell Adhesion Array Kit can be used for assessing the presence or absence of specific integrins on the cell surface. Less<<
SDB (Sicherheitsdatenblätter), Analysenzertifikate und Qualitätszertifikate, Dossiers, Broschüren und andere verfügbare Dokumente.
Cell adhesion plays a major role in cellular communication and regulation, and is of fundamental importance in the development and maintenance of tissues. A family of cell surface receptors, the integrins, mediates cell attachment to proteins of the extracellular matrix. Integrins are heterodimer molecules containing an alpha and beta transmembrane glycoprotein subunit that are non-covalently bound together (1-2). Historically, antibodies have been used to determine the integrin profiles of cells by immunoprecipitation, immunofluorescence, immunoblotting, and flow cytometry. These methods are time-consuming, laborious or require sophisticated equipment.
The CHEMICON® Alpha/Beta Integrin-Mediated Cell Adhesion Array Combo Kit is designed as a cost effective, efficient method for identification of cell surface integrins. The kit format uses 12 x 8-well removable strips in a plate frame for convenience and flexibility in designing assays. The Alpha/Beta Integrin-Mediated Cell Adhesion Arrays can be completed within 1-2 hours, providing a rapid, sensitive, and quantitative assay for the characterization of alpha and beta integrin surface expression.
In addition, Chemicon continues to provide numerous migration, invasion, and adhesion products including:
24-well Insert Cell Migration and Invasion assay systems
CytoMatrix™ Cell Adhesion strips (ECM protein coated)
QuantiMatrix™ ECM protein ELISA Kits
Test Principle
The CHEMICON® Alpha/Beta Integrin-Mediated Cell Adhesion Array Kit uses mouse monoclonal antibodies generated against human alpha (alpha1, alpha2, alpha3, alpha4, alpha5, alphaV, and alphavbeta3), and beta (beta1, beta2, beta3, beta4, beta6, alphaVbeta5, and alpha5beta1) integrins/subunits, that are immobilized onto a goat anti-mouse antibody coated microtiter plate. The plate is then used to capture cells expressing these integrins on their cell surface; goat anti-mouse antibody coated wells are provided as a negative assay control. Each strip (8 wells total) consists of 7 integrin antibody coated wells and one negative control well (see plate layout on page 4). Experimental cells are seeded onto the coated surface and incubated. Subsequently, unbound cells are washed away, and the adherent cells are lysed and detected by the patented CyQuant GR dye (Molecular Probes) (3-4). This green-fluorescent dye exhibits strong fluorescence enhancement when bound to cellular nucleic acids (5). Relative cell attachment is determined using a fluorescence plate reader.
Additional information about the monoclonals provided in the kit: Monoclonal / Known Species Reactivity / Clone Number Mouse anti-Alpha1 [CD49a] / Hu / FB12 Mouse anti-Alpha2 [CD49b] / Hu / P1E6 Mouse anti-Alpha3 [CD49c] / Hu / P1B5 Mouse anti-Alpha4 [CD49d] / Hu, Prim / P1H4 Mouse anti-Alpha5 [CD49e] / Hu / P1D6 Mouse anti-AlphaV [CD51] / Hu / P3G8 Mouse anti-AlphaV/Beta 3 [CD51/CD61] / Hu, Bov, Por, Av, Can, Chk, Mky, Rb / LM609 Mouse anti-Beta1 [CD29] / Hu, Mky, Rb / P4G11 Mouse anti-Beta2 [CD18] / Hu / P4H9 Mouse anti-Beta3 [CD61] / Hu / 25E11 Mouse anti-Beta4 [CD104] / Hu / ASC-9 Mouse anti-Beta6 / Hu, Rt, Mky, Rb / CSb6 Mouse anti-AlphaV/ Beta5 [CD51/CD61] / Hu, Chk / P1F6 Mouse anti-Alpha5/ Beta1 [VLA5, FNR] / Hu, Por / HA5
Materials Required but Not Delivered
· Multi-channel and/or repeating pipettes
· 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.
· Tissue culture growth medium appropriate for subject cells, such as DMEM containing 10% FBS
· Quenching Medium: serum-free medium, such as DMEM, EMEM, 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.
· Sterile PBS or HBSS to wash cells
· Deionized water
· Low speed centrifuge and tubes for cell harvesting
· CO2 incubator appropriate for subject cells
· Sterile cell culture hood
· Hemacytometer and microscope for counting cells
· Trypan Blue or equivalent viability stain
· Fluorescence plate reader with 485nm and 530nm filter set
· 96-well plate suitable for fluorescence measurement
References
Product Information
Components
Alpha Integrin Array Plate: (PN: 90599) One 96-well plate with 12 strips. Each strip contains one well each of the mouse anti-alpha integrin monoclonal antibodies: (alpha1, alpha2, alpha3, alpha4, alpha5, alphaV, alphavbeta3), and one goat anti-mouse negative well. See the Alpha Integrin plate layout on the insert data sheet.
Beta Integrin Array Plate: (PN: 90600) One 96-well plate with 12 strips. Each strip contains one well each of the mouse anti-beta integrin monoclonal antibodies: (beta1, beta2, beta3, beta4, beta6, alphaVbeta5, and alpha5beta1), and one goat anti-mouse negative well. See the Beta Integrin plate layout on the insert data sheet.
4X Cell Lysis Buffer: (Part No. 90130) One bottle - 16 mL
CyQuant GR Dye®: (Part No. 90132) Two vials - 75 μL each
Assay Buffer: (Part No. 90601) Two bottles - 100 mL each
The Alpha/Beta Integrin-Mediated Cell Adhesion Array Kit can be used for assessing the presence or absence of specific integrins on the cell surface.
Application Notes
The CHEMICON® Alpha/Beta Integrin-Mediated Cell Adhesion Array Kit can be used for assessing the presence or absence of specific integrins on the cell surface. This assay can substitute for FACS analysis (6) and is useful for screening human adherent cells for cell surface integrins or for monitoring in vitro cell differentiation or genetic modification of cells.
For Research Use Only. Not for use in diagnostic procedures.
This gene encodes the alpha 1 subunit of integrin receptors. This protein heterodimerizes with the beta 1 subunit to form a cell-surface receptor for collagen and laminin. The heterodimeric receptor is involved in cell-cell adhesion and may play a role in inflammation and fibrosis. The alpha 1 subunit contains an inserted (I) von Willebrand factor type I domain which is thought to be involved in collagen binding.
FUNCTION: SwissProt: P56199 # Integrin alpha-1/beta-1 is a receptor for laminin and collagen. It recognizes the proline-hydroxylated sequence G-F-P-G- E-R in collagen. SIZE: 1179 amino acids; 130848 Da SUBUNIT: Heterodimer of an alpha and a beta subunit. Alpha-1 associates with beta-1. SUBCELLULAR LOCATION: Membrane; Single-pass type I membrane protein. DOMAIN: SwissProt: P56199 The integrin I-domain (insert) is a VWFA domain. Integrins with I-domains do not undergo protease cleavage. SIMILARITY: Belongs to the integrin alpha chain family. & Contains 7 FG-GAP repeats. & Contains 1 VWFA domain.
Physicochemical Information
Dimensions
Materials Information
Toxicological Information
Safety Information according to GHS
Safety Information
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
The Alpha and Beta Integrin Array Plates can be stored at 2° to 8°C in the foil pouch up to their expiration date. Unused strips may be placed back in the pouch for storage. Ensure that the desiccant remains in the pouch, and that the pouch is securely closed. Keep the remaining kit components at 2° to 8°C.
Precautions
No data is available on the biological toxicity of CyQuant GR® dye. This reagent binds to nucleic acids, and as such, should be treated as a potential mutagen which may cause cancer and heritable genetic damage. Handle with caution. The DMSO stock solution should be handled with special caution as DMSO can facilitate the entry of organic molecules into tissues.
Expression of matrix macromolecules and functional properties of breast cancer cells are modulated by the bisphosphonate zoledronic acid. P G Dedes,Ch Gialeli,A I Tsonis,I Kanakis,A D Theocharis,D Kletsas,G N Tzanakakis,N K Karamanos Biochimica et biophysica acta
1820
2011
The extracellular matrix (ECM) components play key roles in the multistep process of cancer growth and progression. Preclinical and clinical data show that bisphosphonates (BPs) may exert direct or indirect antitumoral effects. Despite proven efficiency in cancer treatment, the mechanism by which BPs can interfere with cancer progression remains elusive.
Millipore offers a significant portfolio of well-published, quantitative and optimized live cell, whole-cell, and cell-based activity assays. Study Apoptosis, Angiogensis, Adhesion and more. Weitere Informationen >>
Cell Adhesion Assays
Millipore offers a significant portfolio of well-published, quantitative and optimized live cell, whole-cell, and cell-based adhesion assays. Weitere Informationen >>
