• 我要登录|
  • 免费注册
    |
  • 我的丁香通
    • 企业机构：
    • 成为企业机构
    • 个人用户：
    • 个人中心
  • 移动端
    移动端
丁香通 logo丁香实验_LOGO
搜实验

    大家都在搜

      大家都在搜

        0 人通过求购买到了急需的产品
        免费发布求购
        发布求购
        点赞
        收藏
        wx-share
        分享

        Growing Cells in Geltrex Reduced Growth Factor Basement Membrane Matrix

        互联网

        1330

        实验步骤

         

        There are many applications for Basement Membrane Matrix which require different thicknesses and concentrations. In general, a protein concentration > 9 mg/ml, is used for differentiation studies of primary cells. Extract diluted below 9 mg/ml does not form a gel, and will only support the propagation of primary cells, but not their differentiation. For applications such as endothelial cell differentiation into capillary-like structures (Tube Assay) a thin gel is needed. For applications such as the differentiation of rat aorta tissue into capillary-like structures (Ring Assay), or cell invasion assays, a thick gel is needed. Some applications, such as propagation of primary cells, only need a protein layer and not a protein matrix; therefore, the layer method should be used.

         

        1.        Thin Gel Method:

        1)        Thaw Geltrex. See Precautions.

        2)        Mix Geltrex by slowly pipetting solution up and down; be careful not to introduce air bubbles.

        3)        Place 50 µl per cm2 onto the growth surface.

        4)        Place coated object at 37°C for 30 minutes.

        5)        Coated objects are ready for use.

         

        2.        Thick Gel Method:

        1)        Thaw Geltrex. See Precautions.

        2)        Mix Geltrex by slowly pipetting solution up and down; be careful not to introduce air bubbles.

        3)        Place 150-200 µl per cm2 onto the growth surface.

        4)        Place coated object at 37°C for 30 minutes.

        5)        Coated objects are ready for use

         

        3.        Thin Layer Method (non-gelling):

        1)        Thaw Geltrex. See Precautions.

        2)        Mix Geltrex by slowly pipetting solution up and down; be careful not to introduce air bubbles.

        3)        Dilute the extract to desired concentration in cold serum-free medium. Empirical determination of the optimal coating concentration for your application may be required. A protein concentration of 0.1 mg/ml is a recommended starting concentration for the propagation of primary cells.

        4)        Add a sufficient amount of solution to cover the entire area onto growth surface.

        5)        Place coated object at 37°C for 60 minutes or until dry.

        6)        Coated objects are ready for use.

         

        4.        Thin Gel Method (non-gelling) for hESC Applications:

        1)        Thaw Geltrex. See Precautions.

        2)        Mix Geltrex by slowly pipetting solution up and down; be careful not to introduce air bubbles.

        3)        Dilute 1 ml of Geltrex into 29 ml DMEM/F12 medium (SKU: 10565). Empirical determination of the optimal coating concentration for your application may be required. Volumes can be adjusted accordingly.

        4)        Add a sufficient amount of diluted Geltrex solution to cover the entire area onto growth surface (1.5 ml for 35 mm dish, 3 ml for 60 mm dish).

        5)        Coat the dish and place at room temperature for a minimum of 60 minutes.

        6)        For long term storage, seal each dish with parafilm to minimize dehydration of the Geltrex solution. The coated dish is stable for two weeks when stored at 2 to 8°C. Do not allow coated surface to dry out and maintain a storage temperature of 2 to 8°C to avoid premature gelling.

        7)        At time of use, aspirate Geltrex™ coating and immediately plate cells in pre-equilibrated cell culture medium.

        ad image
        ad image

        相关产品推荐

        GELTREX FLEX LDEV-Free减生长因子hESC验证版, 10mL

        GELTREX FLEX LDEV-Free减生长因子hESC验证版, 10mL

        询价

        HGF/Hepatocyte Growth Factor 兔单抗

        HGF/Hepatocyte Growth Factor 兔单抗

        ￥2299

        二氢乙锭，38483-26-0，A cell-permeable, chemically-reduced ethidium derivative.，阿拉丁

        二氢乙锭，38483-26-0，A cell-permeable, chemically-reduced ethidium derivative.，阿拉丁

        ￥2127.90

        Glomerular Basement Membrane (GBM; dissociated)，≥80%(SDS-PAGE)，阿拉丁

        Glomerular Basement Membrane (GBM; dissociated)，≥80%(SDS-PAGE)，阿拉丁

        ￥1900.90

        LoxP-STOP-loxP-mCherry HeLa Cells，阿拉丁

        LoxP-STOP-loxP-mCherry HeLa Cells，阿拉丁

        ￥5997.90

        相关问答

        问

        plasma membrane proteins是指什么？它位于何处？

        4 回答 639 围观
        2023-06-08

        问

        Stem cells and development投稿

        3 回答 543 围观
        2023-04-27

        问

        SEER数据库无法生成Matrix

        2 回答 367 围观
        2022-10-18

        相关方法

        非限制性体干细胞（unrestricted somatic stem cells，USSCs)的分离

        推荐阅读

        底膜 basement membrane

        Semiquantitative immunoblots of membrane protein-epidermal growth factor

        基膜(basal lamina，basement membrane)

        无忧采购 轻松科研

        无忧采购 轻松科研

        提问
        扫一扫
        丁香实验小程序二维码
        实验小助手
        丁香实验公众号二维码
        扫码领资料
        反馈
        TOP
        打开小程序