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PKH67GL

Kit de lieur cellulaire à fluorescence verte au PKH67 pour le marquage des membranes cellulaires à usage général

Distributed for Phanos Technologies

Synonyme(s) :

Green PKH membrane labeling kit

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A propos de cet article

NACRES:
NA.32
UNSPSC Code:
12352207
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Quality Segment

packaging

pkg of 1 kit

storage condition

protect from light

fluorescence

λex 490 nm; λem 502 nm (PKH67 dye)

detection method

fluorometric

shipped in

ambient

storage temp.

room temp

Application

Ce kit est destiné au marquage des membranes cellulaires à usage général. Le PKH67 possède une queue de carbone aliphatique plus longue que le PKH1 et le PKH2, deux autres colorants verts précédemment décrits pour le suivi cellulaire in vitro et in vivo. Sur la base de la plus grande longueur de la queue, des études internes ont constamment montré une réduction du transfert cellule-cellule pour le PKH67 par rapport au PKH2.
Une perte lente de fluorescence a été observée dans des études in vivo utilisant le PKH1 et le PKH2. Le PKH67 peut présenter des propriétés similaires puisque ce comportement semble être caractéristique des colorants de liaison des cellules vertes, mais pas des colorants de liaison des cellules rouges. La corrélation entre la rétention in vitro de la membrane cellulaire et la demi-vie de fluorescence in vivo dans des cellules non en division permet de prévoir une demi-vie de fluorescence in vivo de 10-12 jours pour la PKH67. D'autres colorants verts de liaison cellulaire ayant une demi-vie similaire ont été utilisés pour suivre le trafic des lymphocytes et des macrophages in vivo sur des périodes de 1 à 2 mois, ce qui suggère que le PKH67 sera également utile pour des études de suivi in vivo de durée moyenne.
Le kit de marquage cellulaire en fluorescence verte PKH67 pour le marquage universel des membranes cellulaires a été utilisé dans les applications suivantes :
  • marquage et analyse des exosomes expulsés par les cellules dans le cancer du sein triple négatif
  • marquage des cellules apoptotiques afin d'étudier le rôle joué par le récepteur de l'ανβ5 dans la liaison et l'internalisation des cellules apoptotiques
  • imagerie en fluorescence[1]

Other Notes

Pour obtenir des détails techniques supplémentaires sur les colorants fluorescents PKH et CellVue® destinés au marquage cellulaire, notamment une bibliographie complète, rendez-vous ici.

Legal Information

CellVue is a registered trademark of Phanos Technologies

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Cet article
MINI67PKH26GLMINI26
fluorescence

λex 490 nm; λem 502 nm (PKH67 dye)

fluorescence

λex 490 nm; λem 502 nm (PKH67 dye)

fluorescence

λex 551 nm; λem 567 nm (PKH26 dye)

fluorescence

λex 551 nm; λem 567 nm (PKH26 dye)

storage condition

protect from light

storage condition

protect from light

storage condition

protect from light

storage condition

protect from light

storage temp.

room temp

storage temp.

room temp

storage temp.

room temp

storage temp.

room temp

shipped in

ambient

shipped in

ambient

shipped in

ambient

shipped in

ambient

packaging

pkg of 1 kit

packaging

pkg of 1 kit

packaging

pkg of 1 kit

packaging

pkg of 1 kit

detection method

fluorometric

detection method

fluorometric

detection method

fluorometric

detection method

fluorometric


Composants de kit seuls

Réf. du produit
Description

  • Diluent C 6 x 10

  • PKH67 Cell Linker in ethanol .5 mL

pictograms

FlameExclamation mark

signalword

Danger

hcodes

Hazard Classifications

Eye Irrit. 2 - Flam. Liq. 2

Classe de stockage

3 - Flammable liquids

flash_point_f

57.2 °F - closed cup

flash_point_c

14 °C - closed cup



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Questions

1–10 of 12 Questions  
  1. I mark mitochondria for injection with PKH26 Red Fluorescent Cell Linker Kit for General Cell Membrane Labeling. I want to use another dye to mark another type of mitochondria and inject them together in the same fish egg. Is PKH67 ok for this like PKH26?

    1 answer
    1. Yes, Product PKH67 would be suitable for this application. In this paper, both Products PKH26 and PKH67 are used in a similar application:
      https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5679712/

      Helpful?

  2. What can be the alternative for removing excess dye after labeling of exosomes using PKH67 dye apart from ultracentrifugation?

    1 answer
    1. Excess dye is bound to serum or albumin which is added to the cell/dye mixture as a stop reaction. The recommended clean up or dye removal is by standard centrifugation (400 x g for 10 minutes, wash, and repeat). See the Product Information Sheet, page 3, bullet point 8 for more information:
      https://www.sigmaaldrich.com/deepweb/assets/sigmaaldrich/product/documents/984/984/pkh67glbul.pdf

      Alternate methods for excess dye removal have not been investigated.

      Helpful?

  3. Can we remove the excess PKH67 DYE used for the labeling of exosomes using exosome spin column rather than ultracentrifugation.

    1 answer
    1. The recommended exosome labeling protocol has been optimized to assure the best possible results. The use of exosome spin columns with the PKH and CellVue products has not been validated. See below for a link to our exosome labeling protocol. A spin column method may not offer the best results.

      https://www.sigmaaldrich.com/technical-documents/protocol/cell-culture-and-cell-culture-analysis/imaging-analysis-and-live-cell-imaging/exosome-labeling-pkh

      Helpful?

  4. What method of fixation can be used for tissue/cells with the PKH Fluorescent Cell Linker Kit for General Cell Membrane Labeling dyes?

    1 answer
    1. A protocol for visualization of tissue sections can be found in the technical bulletin.  For visualization of stained cells by immunofluorescence or flow cytometry, the cells can be fixed in 2% paraformaldehyde for 15 minutes. The use of other organic solvents will extract the dye from the cells.  If internal labeling is desired, the cells can be permeabilized with saponin (50-75 μg/mL). Here is the link to the Troubleshooting Guide.

      Helpful?

  5. Are cells lost during the PKH67 Green Fluorescent Cell Linker Kit for General Cell Membrane Labeling staining process?

    1 answer
    1. Over-labeling of the cells will result in loss of membrane integrity and reduced cell recovery.  Methods for improving cell viability can be found on the troubleshooting guide.

      Helpful?

  6. Does the dye leak from the cells after labeling when using Product PKH67GL, PKH67 Green Fluorescent Cell Linker Kit for General Cell Membrane Labeling?

    1 answer
    1. In general, slow loss of fluorescence has been observed with PKH1 and PKH2 in in vivo studies and it is likely that PKH67 may also slowly lose fluorescence. Leakage (or loss of staining intensity) may also occur due to cell-to-cell transfer of the dye. The most common cause of cell-to-cell dye transfer is inadequate washing of the cells. Wash cells 3-5 times after labeling and transfer samples to new tubes between washes.

      Helpful?

  7. What is the excitation and emission spectra for Product PKH67GL, Green Fluorescent Cell Linker Kit?

    1 answer
    1. The spectra can be found on the product insert.The product has a maximum excitation at 490 nm and maximum emission at 502 nm.

      Helpful?

  8. How many cells can be stained with Product PKH67GL, Green Fluorescent Cell Linker Kit?

    1 answer
    1. This kit can stain 50 × 107 cells if used as directed (1 × 107 cells stained with 2 × 10-6 M PKH67).

      Helpful?

  9. What is the Department of Transportation shipping information for this product?

    1 answer
    1. Transportation information can be found in Section 14 of the product's (M)SDS.To access the shipping information for this material, use the link on the product detail page for the product.

      Helpful?

  10. When using Product PKH67GL, PKH67 Green Fluorescent Cell Linker Kit for General Cell Membrane Labeling, how long will the cells remain stained after treatment?

    1 answer
    1. Labeled cells that have been washed can be visualized in culture up to 100 days after staining (for non-dividing cells). The dye itself is stable and will divide equally when the cells divide. After staining with PKH dyes, you can observe as many as 8 divisions depending on how brightly the cells were stained initially and the amount of surface area on the cells. Most commonly, 4-6 divisions can be visualized.

      Helpful?

1–10 of 12 Questions  

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