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Phalloidin Control (21834) in ICC/IF
Immunofluorescent analysis of Calreticulin (green) 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 1:50 for at least 1 hour at room temperatu... View More
Product Details
21834
Applications
Tested Dilution
Immunohistochemistry (IHC)
Assay-dependent
Immunocytochemistry (ICC/IF)
1-5 units/mL
Product Specifications
Type
Control
Conjugate
Form
Lyophilized
Concentration
300 U/mL
Storage conditions
-20°C
Shipping conditions
Wet ice
RRID
AB_2532156
Product Specific Information
Format: 300 units of lyophilized DyLight 554-Phalloidin (300 units/mL) in methanol. Prepare stock solution by adding 1000 µL of pure methanol to the vial and gently mix.
21834 has been successfully used in immunofluorescence and immunohistochemistry. DyLight 554-Phalloidin has an excitation/emission of 551/572 nm and molecular weight of 1313.5 g/mole.
DyLight 554-Phalloidin Stock Solution can be prepared in methanol, DMF, or DMSO and unused Stock Solution should be promptly stored at -20°C to minimize evaporation. Working solutions (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 antibodies or dyes before using DyLight 554-Phalloidin.
Target Information
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.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
Performance Guarantee
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