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Phalloidin Control (21833) in ICC/IF
Immunofluorescent analysis of Phalloidin (green) and Calreticulin (red) in U2OS cells. Formalin fixed cells were permeabilized with 0.1% Triton X-100 in PBS for 10 minutes at room temperature and blocked with 2% BSA (Product # 37525) in PBS + 0.1% Triton X-100 for 30 minutes at room temperature. Cells were probed with a Calreticulin polyclonal antibody (Product # PA3-900) at a dilution of 1:75 for at least ... View More
产品信息
21833
应用
建议稀释比
免疫组化 (IHC)
Assay-dependent
免疫细胞化学 (ICC/IF)
1:100-1:1,000
产品规格
类型
Control
偶联物
激发/发射光谱
492/519 nm
查看光谱
形式
Lyophilized
浓度
300 U/mL
保存条件
-20°C
运输条件
Ambient
RRID
AB_2532155
产品详细信息
Format: 300 units of lyophilized DyLight 488-Phalloidin (300 units/mL) in methanol. Prepare stock solution by adding 1000 µL of pure methanol to the vial and gently mix.
21833 has been successfully used in immunofluorescence and immunohistochemistry. DyLight 488-Phalloidin has an excitation/emission of 495/521 nm and molecular weight of 1582.65 g/mole.
DyLight 488-Phalloidin Stock Solution is prepared in methanol, and is also soluble in DMF, or DMSO. The unused Stock Solution should be promptly stored at -20°C in a foil pouch with desiccant, and protected from light. The stock solutions are stable for at least one year. Staining cells with some of the DyLight Phalloidin conjugates may require the use of a higher concentration of the phalloidin conjugate; therefore to minimize the amount of methanol added to the cells, the vial contents could be dissolved in 0.5 mL methanol to yield a final concentration of 600units/mL.
Working solution of 1unit/mL (per 96-well plate) can be prepared by diluting 20 µL of the Stock Solution in 5.98 mL of PBS and mixing well. Typical staining procedure adds 50 µL of Working Solution (i.e. 1 to 5 unit/mL) to a each well. Incubate cells in the dark for 30 minutes at room temperature (optimal staining times varies from 10 minutes to 3 hours depending on cell type). Aspirate and wash cells three times in PBS after incubation. If desired, probe with specific primary antibodies followed by secondary antibodies conjugated to any compatible fluorophores before using DyLight 488-Phalloidin conjugate.
靶标信息
Phalloidin is a bicyclic peptide that belongs to a family of toxins isolated from the deadly Amanita phalloides “death cap” mushroom and is commonly used as a counterstain (similar to DAPI or Hoechst) in cell biology and histology imaging applications to selectively label F-actin in fixed cells, permeabilized cells, and cell-free experiments. Labeled phalloidin conjugates have similar affinity for both large and small filaments and bind in a stoichiometric ratio of about one phallotoxin per actin subunit in both muscle and non-muscle cells. Phalloidins reportedly do not bind to monomeric G-actin, unlike some antibodies against actin. The dynamics of the actin polymerization in cells are important for a variety of cellular processes from cell motility to cell shape, from muscular contraction to cytokinesis, and more.
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