Key Specifications Table
| Key Applications | Detection Methods |
|---|---|
| ACT | Chromogenic |
| Description | |
|---|---|
| Catalogue Number | ECM102 |
| Brand Family | Chemicon® |
| Trade Name |
|
| Description | Millicoat™ Human Vitronectin 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 Vitronectin. 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 |
|
| Product Information | |
|---|---|
| Detection method | Chromogenic |
| Quality Level | MQ100 |
| Applications | |
|---|---|
| Application | Millicoat Cell Adhesion Strips are provided as 12 x 8-well removable strips in a 96-well plate frame for convenience & flexibility in designing assays. |
| Key Applications |
|
| 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 45 minutes 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 strips 3-5 times with PBS (300 mL/well) to remove the excess stain. 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 | |
|---|---|
| Species Reactivity |
|
| Entrez Gene Number |
|
| Entrez Gene Summary | The protein encoded by this gene is a member of the pexin family. It is found in serum and tissues and promotes cell adhesion and spreading, inhibits the membrane-damaging effect of the terminal cytolytic complement pathway, and binds to several serpin serine protease inhibitors. It is a secreted protein and exists in either a single chain form or a clipped, two chain form held together by a disulfide bond. |
| Gene Symbol |
|
| UniProt Number |
|
| UniProt Summary | FUNCTION: SwissProt: P04004 # Somatomedin-B is a growth hormone-dependent serum factor with protease-inhibiting activity. SIZE: 478 amino acids; 54306 Da SUBUNIT: Exists in two forms: a single chain 75 kDa form (V75) and a clipped form composed of two chains (65 kDa and 10 kDa) (V65+V10) which are held together by a disulfide bond. Interacts with SERPINE1/PAI1 and insulin. SUBCELLULAR LOCATION: Secreted, extracellular space. TISSUE SPECIFICITY: Plasma. DOMAIN: SwissProt: P04004 The SMB domain mediates interaction with SERPINE1/PAI1. The heparin-binding domain mediates interaction with insulin. PTM: Sulfated on 2 tyrosine residues. & N- and O-glycosylated (By similarity). & Phosphorylation on Thr-69 and Thr-76 favors cell adhesion and spreading. & It has been suggested that the active SMB domain may be permitted considerable disulfide bond heterogeneity or variability, thus two alternate disulfide patterns based on 3D structures are described with 1 disulfide bond conserved in both.SIMILARITY: Contains 4 hemopexin-like domains. & Contains 1 SMB (somatomedin-B) domain. |
| Stem Cell Type |
|
| Product Usage Statements | |
|---|---|
| Usage Statement |
|
| Storage and Shipping Information | |
|---|---|
| Storage Conditions | Strips 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 |