Phalloidin CruzFluor™ 647 Conjugate MW: ~2200

Phalloidin CruzFluor™ 647 Conjugate

Phalloidin CruzFluor 647 Conjugate is rated 5.0 out of 5 by 1.
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Application: Phalloidin CruzFluor™ 647 Conjugate is an F-actin specific fluorescent phalloidin conjugate
Molecular Weight: ~2200
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™ 647 Conjugate is a deep red fluorescent conjugate that is an excellent substitute for AlexFluor® 647. Phalloidin is a heptapeptide phallotoxin produced by Amanita phalloides, the death cap mushroom. Phalloidin 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 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.

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 :
≤ 100% phalloidin
Appearance :
Powder
Physical State :
Solid
Solubility :
Soluble in DMSO.
Storage :
Desiccate at -20° C
Fluoresence :
λex 650 nm, λem 665 nm
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.

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Certificate of Analysis

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Phalloidin CruzFluor™ 647 Conjugate  Product Citations

See how others have used Phalloidin CruzFluor™ 647 Conjugate. Click on the entry to view the PubMed entry .

Citations 1 to 9 of 9 total

PMID: # 33824207  Diggins, NL.|Crawford, LB.|Hancock, MH.|Mitchell, J.|Nelson, JA.| et al. 2021. mBio. 12:

PMID: # 33571402  Zeigler, AC.|Chandrabhatla, AS.|Christiansen, SL.|Nelson, AR.|Holmes, JW.|Saucerman, JJ.| et al. 2021. CPT Pharmacometrics Syst Pharmacol.

PMID: # 28528863  Inostroza-Brito, KE.|Collin, EC.|Majkowska, A.|Elsharkawy, S.|Rice, A.|Del Río Hernández, AE.|Xiao, X.|Rodríguez-Cabello, J.|Mata, A.| et al. 2017. Acta Biomater. 58: 80-89.

PMID: # 28065854  Sušjan, P.|Roškar, S.|Hafner-Bratkovič, I.| et al. 2017. Biochem. Biophys. Res. Commun. 483: 823-828.

PMID: # 26438793  Carrasco, SE. et al. 2015. Infect. Immun. 83: 4848-60.

PMID: # 24623484  Marques, JG. et al. 2014. Pharm. Res. -.

PMID: # 24434137  Dev Cell. 70-80.

PMID: # 31920691  Front Physiol. 1481.

PMID: # 28767724  PLoS One. e0181579.

Citations 1 to 9 of 9 total

Hi, can Phalloidin CruzFluor™ 647 Conjugate be be used with antibodies when performing cytoimmunofluorescence double or triple stainings. If yes, so how to use it? During incubation with secondary antibody? Do you have a protocol or any suggetions?

Asked by: Zina
Thank you for your question. Yes, our Phalloidin CruzFluor conjugates can be used alongside antibodies in IF staining of formalin-fixed cells and formalin-fixed, frozen tissue sections. The Phalloidins will stain F-actin. Paraffin-embedded tissues are not recommended for Phalloidin staining, due to the use of solvents during the deparaffinization process that will disrupt actin. Please contact our Technical Service for a protocol.
Answered by: Technical Support
Date published: 2018-12-18

How much DMSO is needed to solubilize for 300 tests?

Asked by: SCchem16
Thank you for your question. For this chemical, sc-363797: Phalloidin CruzFluor™ 647 Conjugate, a 1000 X Phalloidin CruzFluor DMSO stock solution can be prepared by adding 30 uL of DMSO into the powder form vials.
Answered by: Chemical Support 2
Date published: 2017-03-11

What are the storage recommendations for the solubilized product?

Asked by: SCchem16
Thank you for your question. This chemical, Phalloidin CruzFluor™ 647 Conjugate, once in solution should be aliquoted and stored at -20ºC and protected from light.
Answered by: Chemical Support 2
Date published: 2018-10-06
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Rated 5 out of 5 by from Various publications cite the use of Phalloidin Various publications cite the use of Phalloidin CruzFluor 647 Conjugate 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. -SCBT Publication Review
Date published: 2015-02-25
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