Vimentin-Label (Atto488)

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Protein- /Substanz
  • CTRCT30
  • cb28
  • vime
  • vim
  • vim1
  • vim2
  • VIM
  • Vimentin
  • vimentin
  • vimentin L homeolog
  • VIM
  • Vim
  • vim
  • vim.L
Reaktivität
Human, Säugetier
Konjugat
Atto 488
Applikation
Fluorescence Microscopy (FM), Super-resolution Microscopy (SRM), Confocal Microscopy (CM), Immunofluorescence (IF), Wide-field Fluorescence Microscopy (WFFM)
Optionen
Hersteller
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Hersteller Produkt- Nr.
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Verwendungszweck With our Booster you reactivate, boost, stabilizate the signals of your fusion proteins.
Spezifität Vimentin-Booster specifically binds to the vimentin intermediate filament protein. Validated on MDCK cells and on human A549 cells upon stimulation with TGFβ.
Produktmerkmale Vimentin-Booster for immunostaining of vimentin intermediate filaments in fixed cells. Vimentin-Booster consists of a vimentin-specific alpaca antibody domain conjugated to the fluorescent dye ATTO 488
Reinigung Purified
Andere Bezeichnung Vimentin
Hintergrund The vimentin filament network plays a key role in cell architecture and signaling in mesenchymal cells. Vimentin is also an important marker of the epithelial-mesenchymal transition, a highly dynamic cellular process involved in the initiation of metastasis and cancer progression.
Applikationshinweise For immunofluorescence of vimentin intermediate filaments in fixed cells
ATTO488:
Excitation range 480 - 510 nm (λabs= 501 nm)
Emission range 520 - 560 nm (λfl= 523 nm)
IF/ICC: 1/200 - 1/400

Optimal working concentration is application-dependent and should be determined by testing a range of dilutions from 1/100 to 1/1000.
Kommentare

Vimentin-Booster is a small vimentin-binding protein derived from a camelid VHH / socalled nanobody and conjugated to the fluorescent dye ATTO 488 (from ATTO-TEC). Immunofluorescence staining with alpaca Nano-Boosters is compatible with different fixation protocols and does not require any secondary antibody. Due to their small size, alpaca Nano-Boosters show better tissue penetration and improve staining precision, which allows obtaining higher resolution images.
Booster are very small, highly specific GFP- or RFP-binding proteins covalently coupled to the superior fluorescent dyes from ATTO-TEC.

Testdurchführung
  • 1. Fixation: Wash cells seeded on coverslips twice with PBS (Phosphate Buffered Saline). Fix in 3.7 % formaldehyde in PBS for 10 min at room temperature.
    Note: Always prepare a fresh formaldehyde dilution.
    Note: Alternatively, it is possible to use methanol or acetone:methanol for fixation: apply ice-cold 100 % methanol or 1:1 acetone:methanol onto cells for 5 min.
  • 2. Wash samples three times with PBS. Do not store fixed cells.
  • 3. Permeabilization and blocking: Add PBS containing 0.2 % Triton X-100 and 3 % BSA to samples and incubate for 1 h at room temperature.
  • 4. Wash samples three times with PBS.
  • 5. Vimentin-Booster incubation: Dilute Vimentin-Booster 1:200 in PBS supplemented with 3 % BSA. Apply onto samples and incubate overnight at 4 °C.
    Note: For multiplexing protocols, you can combine Vimentin-Booster with other primary or secondary antibodies.
  • 6. Wash samples three times for 5-10 min in PBS.
  • 7. If required, counterstain with DNA fluorescent dyes, e.g. DAPI in PBS. Wash samples with PBS. Proceed with imaging of the samples in PBS or imaging buffer.
  • 8. Post-fixation and mounting: Alternatively, if mounting of samples is required, postfix samples with 3.7 % formaldehyde in PBS for 10 min at room temperature. Wash samples three times with PBS and mount in a medium of choice (Mowiol®, VECTASHIELD®, ProLong® Diamond Antifade Mountant, or other).

  • Suggested buffer composition:
  • Fixation buffer: 3.7 % formaldehyd in PBS
  • Permeabilization and blocking buffer: 0.2 % Triton X-100 (w/w), 3 % BSA (w/v), PBS
  • Wash buffer: PBS
  • Immunostaining buffer: 3 % BSA (w/v), PBS
Beschränkungen Nur für Forschungszwecke einsetzbar
Format Liquid
Konzentration 0.3-0.5 mg/mL
Buffer Purified recombinant protein in 1x PBS, 0.09 % sodium azide
Konservierungsmittel Sodium azide
Vorsichtsmaßnahmen This product contains Sodium azide: a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.
Handhabung Do not freeze. Protect from light.
Lagerung 4 °C
Informationen zur Lagerung Shipped at ambient temperature. Upon receipt store at +4°C. Stable for 6 month. Do not freeze. Protect from light.
Haltbarkeit 6 months