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Key Specifications Table
| Key Applications | Detection Methods |
|---|---|
| Enzyme Assays | Chromogenic |
| Description | |
|---|---|
| Catalogue Number | ECM103 |
| Brand Family | Chemicon® |
| Trade Name |
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| Description | Millicoat™ Human Laminin Coated Strips (96-Wells) |
| Overview | Cell adhesion plays a major role in cellular communication and regulation, and is of fundamental importance in the development and maintenance of tissues. Scientists are continually examining the adhesion and migration of many diverse cell types on various extracellular matrix (ECM) component proteins. Millicoat™ Cell Adhesion Strips are provided as 12 x 8-well removable strips in a plate frame for convenience and flexibility in designing assays. The wells in rows A - G have been coated with Human Laminin. Row H of each strip is coated with BSA which serves as a negative assay control. Cells are seeded onto the coated substrate. Subsequently, adherent cells are fixed and stained. Relative attachment is determined using absorbance readings. |
| Alternate Names |
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| Product Information | |
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| Detection method | Chromogenic |
| Quality Level | MQ100 |
| Applications | |
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| Application | 96-well plate coated with human laminin used for cell adhesion studies. |
| Key Applications |
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| Application Notes | PROCEDURE: NOTE: Optimal assay timing and performance may vary for different cell lines but generally can be obtained using subconfluent cell cultures in the assay described below. Subconfluent cultures can be achieved by splitting cells 1 to 2 days prior to performing the assay. 1. Rehydrate the strips with 200 mL of PBS per well for at least 15 minutes at room temperature. Remove the PBS from the rehydrated strips. 2. Prepare a single cell suspension, preferably using a non-enzymatic dissociation buffer. Optimum cell density may be determined by titration of the cells. A common starting range is between 1x10E05 to 1x10E07 cells/mL. 3. Add 100 mL of the diluted cell suspension to each well. Incubate the plate at 37°C for 1 hour in a CO2 incubator. Gently wash the plate 2-3 times with PBS containing Ca2+/Mg2+ (200 mL/well). 4. Add 100 mL of 0.2% crystal violet in 10% ethanol to each well. Incubate for 5 minutes at room temperature. Remove the stain from the wells. Gently wash the plate 3 times with PBS (300 mL/well). 5. Add 100 mL of Solubilization Buffer (A 50/50 mixture of 0.1M NaH2PO4, pH 4.5 and 50% ethanol) to each well. Allow strips to incubate and gently shake at room temperature until the cell-bound stain is completely solubilized; approximately 5 minutes. 6.Determine the absorbance at 540 - 570 nm on a microplate reader. |
| Biological Information | |
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| Species Reactivity |
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| Entrez Gene Number |
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| Entrez Gene Summary | Laminins, a family of extracellular matrix glycoproteins, are the major noncollagenous constituent of basement membranes. They have been implicated in a wide variety of biological processes including cell adhesion, differentiation, migration, signaling, neurite outgrowth and metastasis. Laminins are composed of 3 non identical chains: laminin alpha, beta and gamma (formerly A, B1, and B2, respectively) and they form a cruciform structure consisting of 3 short arms, each formed by a different chain, and a long arm composed of all 3 chains. Each laminin chain is a multidomain protein encoded by a distinct gene. Several isoforms of each chain have been described. Different alpha, beta and gamma chain isomers combine to give rise to different heterotrimeric laminin isoforms which are designated by Arabic numerals in the order of their discovery, i.e. alpha1beta1gamma1 heterotrimer is laminin 1. The biological functions of the different chains and trimer molecules are largely unknown, but some of the chains have been shown to differ with respect to their tissue distribution, presumably reflecting diverse functions in vivo. This gene encodes the beta chain isoform laminin, beta 1. The beta 1 chain has 7 structurally distinct domains which it shares with other beta chain isomers. The C-terminal helical region containing domains I and II are separated by domain alpha, domains III and V contain several EGF-like repeats, and domains IV and VI have a globular conformation. Laminin, beta 1 is expressed in most tissues that produce basement membranes, and is one of the 3 chains constituting laminin 1, the first laminin isolated from Engelbreth-Holm-Swarm (EHS) tumor. A sequence in the beta 1 chain that is involved in cell attachment, chemotaxis, and binding to the laminin receptor was identified and shown to have the capacity to inhibit metastasis. |
| Gene Symbol |
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| UniProt Number |
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| UniProt Summary | FUNCTION: SwissProt: P07942 # Binding to cells via a high affinity receptor, laminin is thought to mediate the attachment, migration and organization of cells into tissues during embryonic development by interacting with other extracellular matrix components. SIZE: 1786 amino acids; 198066 Da SUBUNIT: Laminin is a complex glycoprotein, consisting of three different polypeptide chains (alpha, beta, gamma), which are bound to each other by disulfide bonds into a cross-shaped molecule comprising one long and three short arms with globules at each end. Beta-1 is a subunit of laminin-1 (EHS laminin), laminin-2 (merosin), and laminin-6 (K-laminin). SUBCELLULAR LOCATION: Secreted, extracellular space, extracellular matrix, basement membrane. Note=Major component. DOMAIN: SwissProt: P07942 The alpha-helical domains I and II are thought to interact with other laminin chains to form a coiled coil structure. & Domains VI and IV are globular. SIMILARITY: Contains 13 laminin EGF-like domains. & Contains 1 laminin IV type B domain. & Contains 1 laminin N-terminal domain. |
| Stem Cell Type |
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| Product Usage Statements | |
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| Usage Statement |
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| Storage and Shipping Information | |
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| Storage Conditions | May be stored at 2-8°C in the foil pouch for at least 3 months. 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. |
| Packaging Information | |
|---|---|
| Material Size | 1 plate |
| Material Package | 96 wells |