Vimentin-Label (Atto488)

Details for Product No. ABIN3071449, Supplier: Log in to see
Target Name (Antigen)
  • CTRCT30
  • cb28
  • vime
  • vim
  • vim1
  • vim2
  • VIM
  • Vimentin
  • vimentin
  • vimentin L homeolog
  • VIM
  • Vim
  • vim
  • vim.L
Reactivity
Human, Mammalian
Conjugate
Atto 488
Application
Fluorescence Microscopy (FM), Super-resolution Microscopy (SRM), Confocal Microscopy (CM), Immunofluorescence (IF), Wide-field Fluorescence Microscopy (WFFM)
Options
Supplier
Log in to see
Supplier Product No.
Log in to see
Purpose With our Booster you reactivate, boost, stabilizate the signals of your fusion proteins.
Specificity Vimentin-Booster specifically binds to the vimentin intermediate filament protein. Validated on MDCK cells and on human A549 cells upon stimulation with TGFβ.
Characteristics 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
Purification Purified
Alternative Name Vimentin
Background 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.
Research Area Cytoskeleton, Lineage Markers, Cell/Tissue Markers, Neural Stem Cell marker
Application Notes 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.
Comment

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.

Assay Procedure
  • 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
Restrictions For Research Use only
Format Liquid
Concentration 0.3-0.5 mg/mL
Buffer Purified recombinant protein in 1x PBS, 0.09 % sodium azide
Preservative Sodium azide
Precaution of Use This product contains Sodium azide: a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.
Handling Advice Do not freeze. Protect from light.
Storage 4 °C
Storage Comment Shipped at ambient temperature. Upon receipt store at +4°C. Stable for 6 month. Do not freeze. Protect from light.
Expiry Date 6 months