推荐产品
product name
Millicoat® Human Collagen Type IV Coated Strips (96-Wells), 96-well plate coated with human Collagen Type IV used for cell adhesion studies.
品質等級
物種活性
human
製造商／商標名
Chemicon®
Millicoat®
技術
activity assay: suitable
cell based assay: suitable
輸入
sample type: mouse embryonic stem cell(s)
sample type hematopoietic stem cell(s)
sample type epithelial cells
sample type neural stem cell(s)
sample type induced pluripotent stem cell(s)
sample type pancreatic stem cell(s)
sample type mesenchymal stem cell(s)
sample type: human embryonic stem cell(s)
NCBI登錄號
UniProt登錄號
檢測方法
colorimetric
運輸包裝
wet ice
基因資訊
human ... COL4A1(1282)
一般說明
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 Collagen Type IV. 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.
應用
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.
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.
Research Category
Cell Structure
Cell Structure
包裝
96 wells
儲存和穩定性
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.
法律資訊
CHEMICON is a registered trademark of Merck KGaA, Darmstadt, Germany
MILLICOAT is a registered trademark of Merck KGaA, Darmstadt, Germany
免責聲明
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.
儲存類別代碼
11 - Combustible Solids
水污染物質分類（WGK）
WGK 3
閃點（°F）
Not applicable
閃點（°C）
Not applicable
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实验方案
This page covers the ECM coating protocols developed for four types of ECMs on Millicell®-CM inserts, Collagen Type 1, Fibronectin, Laminin, and Matrigel.
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