Phalloidin CruzFluor™ 555 Conjugate MW: ~1300

Phalloidin CruzFluor™ 555 Conjugate

Phalloidin CruzFluor 555 Conjugate is rated 5.0 out of 5 by 1.
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Alternate Names: Phalloidin CruzFluor™ 555 Conjugate is also known as Phalloidin-iFluor™ 555 Conjugate.
Application: Phalloidin CruzFluor™ 555 Conjugate is a fluorescent phalloidin conjugate that selectively binds to F-actins.
Molecular Weight: ~1300
Supplemental Information: This is classified as a Dangerous Good for transport and may be subject to additional shipping charges.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.
* Refer to Certificate of Analysis for lot specific data (including water content).
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Phalloidin CruzFluor™ 555 Conjugate is an orange fluorescent conjugate that is an excellent substitute for AlexFluor® 555. Phalloidin CruzFluor™ 555 Conjugate is a heptapeptide phallotoxin produced by Amanita phalloides, the death cap mushroom. Phalloidin CruzFluor™ 555 Conjugate binds and stabilizes filamentous F-actin, preventing the dissociation of paired filaments and disturbing the natural equilibrium between filamentous (polymeric) and globular (monomeric) actin in the cell, as well as inhibit the ATP hydrolysis activity of F-actin. Actin is used by the cell for mechanical processes such as growth, scaffolding of the cytoskeleton, and locomotion, and disruption of this actin equilibrium destroys cellular functioning. The magnitude by which the actin protein differs from species to species is no more than 20%.
These properties of Phalloidin CruzFluor™ 555 Conjugate make Phalloidin CruzFluor™ Conjugates a fantastic tool for labeling, identifying and quantitating F-actins. The Phalloidin CruzFluor™ Conjugates are used at nanomolar concentrations with cell-free experiments, cell culture experiments, formaldehyde-fixed, permeabilized tissue sections and the study of actin networks. Because Phalloidin CruzFluor™ Conjugates are offered in different colors the series is excellent for multicolor imaging applications. Phalloidin CruzFluor™ 555 Conjugate is also known as Phalloidin-iFluor™ 555 Conjugate.

Usage :
Phalloidin CruzFluor™ conjugates can be used alongside antibodies in IF staining of formalin-fixed cells and formalin-fixed, frozen tissue sections. The Phalloidin will stain F-actin.

1. Prepare 1000 X Phalloidin DMSO stock solution: by adding 30 uL of DMSO into the powder form vials.

2. Prepare 1X Phalloidin conjugate working solution: by adding 1 uL of 1000X Phalloidin conjugate DMSO solution to 1 mL of PBS with 1% BSA

Note 1: The unused 1000X DMSO stock solution of Phalloidin conjugate should be aliquoted and stored at -20° C. protected from light.

Note 2: Different cell types might be stained differently. The concentration of Phalloidin conjugate working solution should be prepared accordingly.


3. Stain the cells or tissue:

3.1 Perform formaldehyde fixation. Incubate cells with 3.0-4.0% formaldehyde in PBS at room temperature for 10-30 minutes.

Note 3: Avoid any methanol containing fixatives since methanol can disrupt actin during the fixation process. The preferred fixative is methanol-free formaldehyde.

3.2 Rinse the fixed cells 2-3 times in PBS.

3.3 Optional: Add 0.1% Triton X-100 in PBS into fixed cells (from Step 3.2) for 3 to 5 minutes to increase permeability. Rinse the cells 2-3 times in PBS.

3.4 Add Phalloidin conjugate working solution (from Step 2) into the fixed cells (from Step 3.2 or 3.3), and stain the cells at room temperature for 20 to 90 minutes.

3.5 Rinse cells gently with PBS 2-3 times to remove excess Phalloidin conjugate before mounting, sealing and imaging under microscope.

Note 4: If antibody staining is desired, perform blocking, primary antibody and secondary antibody incubations separately between Steps 3.3 and 3.4 according to manufacturer′s protocol. Labeling with Phalloidins can be combined with secondary antibody staining, as long as the correct dilutions of each reagent are used in the staining solution.

Note 5: Paraffin-embedded tissues are not recommended for Phalloidin staining, because the use of solvents during the deparaffinization process will disrupt actin.
Formulation :
1000X stock solution in DMSO; 30 µl total
Physical State :
Liquid
Solubility :
Soluble in DMSO, and water (low).
Storage :
Store at -20° C
Fluoresence :
λex 556 nm, λem 574 nm in DMSO
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.

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Certificate of Analysis

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Phalloidin CruzFluor™ 555 Conjugate  Product Citations

See how others have used Phalloidin CruzFluor™ 555 Conjugate. Click on the entry to view the PubMed entry .

Citations 1 to 6 of 6 total

PMID: # 30746686  2019. J. Cell. Physiol.

PMID: # 29470914  ACS Appl Mater Interfaces. 8507-8518.

PMID: # 32924006  Mater Today Bio. 100071.

PMID: # 33001475  EMBO J. e103530.

PMID: # 32990682  J Clin Invest.

PMID: # 30156819  ACS Appl Mater Interfaces. 31168-31177.

Citations 1 to 6 of 6 total
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Rated 5 out of 5 by from Strong red fluorescence staining was observed Strong red fluorescence staining was observed by IF following incubation of formalin fixed HepG2 cells with a F-actin antibody conjugated to Phalloidin CruzFluor 555 Conjugate. Crisp red fluorescence observed. -SCBT QC
Date published: 2015-01-16
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