Flow cytometry: 20 μL (ready for use). For details see protocol below. Additional information: Clone 53D11 (purified antibody) has been reported by customer to stain Podocalyxin in Formalin-Fixed, Paraffin-Embedded human kidney tissue at an antibody concentration of 10 μg/mL.
实验流程
Flow cytometric analysis for adherent cells We usually use Fisher tubes or equivalents as reaction tubes for all steps after 2). 1) Detach the cells from culture dish by using cell dissociation buffer. 2) Wash the cells 3 times with washing buffer [PBS containing 2 % fetal calf serum (FCS) and 0.1 % NaN 3]. 3) Resuspend the cells with washing buffer (5x10e6 cells/mL). 4) Add 50 μ L of the cell suspension into each tube, and centrifuge at 500 x g for 1 mi nute at room temperature. Remove supernatant by careful aspiration. 5) Add 20 μ L of Clear Back to the cell pellet after tapping. Mix well and incubate for 5 minutes at room temperature. 6) Add 20 μL of the PE labeled anti-Human podocalyxin/PCLP1 monoclonal antibody. Mix well and incubate for 30 minut es at room temperature. 7) Add 1 mL of the washing buffer followed by centrifugation at 500 x g for 1 minute at room temperature. Remove supernatant by careful aspiration. 8) Resuspend the cells with 500 μ L of the washing buffer and analyze by a flow cytometer. (Positive control for Flow cytometry HUVEC)
限制
仅限研究用
状态
Liquid
储存条件
4 °C
储存方法
Store at 2-8 °C. Shelf life: one year from despatch.
有效期
12 months
抗原
PODXL
(Podocalyxin-Like (PODXL))
别名
podocalyxin,podxl
背景
Recent studies with avian embryos and murine embryonic stem cells have suggested that hematopoietic cells are derived from hemangioblasts, the common precursors of hematopo ietic and endothelial cells. Hara et al. molecularly cloned podo calyxin-like protein 1 (PCLP1) as a novel surface marker for endothelial-like cells in the AGM (aorta-gonad-mesonephros) region of mouse embryos, where long-term repopulating hematopoietic stem cells (LTR-HSCs) are known to arise. PCLP1 + CD45 - cells in the AGM region incorporated acetylated low-density lipop rotein and produced both hematopoietic and endothelial cells when cocultured with OP9 stromal cells. Moreover, multiple lineages of hematopoietic cells were generated in vivo when PCLP1 + CD45 - cells were injected in to neonatal liver of busulfan-treated mice. Today it is reported that the PCLP1 is identical with the Podocalyxin.