Vimentin-Label

Description
anti-vimentin VHH/ Nanobody conjugated to the fluorophore ATTO488 for immunofluorescence & microscopy

Conjugations
This primary Nanobody is available as conjugation to ATTO488.

Specificity
Vimentin-Label specifically binds to the vimentin intermediate filament protein. Validated on canine MDCK cells, rodent BHK cells and on human A549 cells upon stimulation with TGFβ.

Applications
Immunofluorescence (IF)
Immunocytochemistry (ICC)
Super Resolution Microscopy (SRM)

Product Size Code Price Buy
Product Vimentin-Label Atto488 Size 10 µL Code vba488-10 Price $ 100
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Product Vimentin-Label Atto488 Size 100 µL Code vba488-100 Price $ 365
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Coupled Nanobody/ VHH
Recombinant, monoclonal, bivalent anti-Vimentin single domain antibody (sdAb) fragment

Specificity
Vimentin-Label specifically binds to the vimentin intermediate filament protein. Validated on canine MDCK cells, rodent BHK cells and on human A549 cells upon stimulation with TGFβ.

Host/isotype
Alpaca/recombinant VHH domain, monoclonal

Available conjugates
ATTO488

Recommended dilution
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 1:1,000.

Microscopy techniques
Wide-field or epifluorescence microscopy; confocal microscopy; super-resolution microscopy e.g. STED, dSTORM

Form
Purified recombinant protein in PBS supplemented with preservative 0.09 % sodium azide

Size
10 μL, 100 μL
Because of the small size of the VHH these volumes correspond to about 10 times the volume of conventional IgG antibodies.

Protein concentration
0.3 – 0.5 mg/mL (conjugates)

Storage instructions
Shipped at ambient temperature. Upon receipt store at 4°C. Do not freeze. Protect from light.

Validation
Validated in cell culture & cell lines and fixed cells: canine MDCK cells, rodent BHK cells and on human A549 cells upon stimulation with TGFβ.

Coupled Nanobody/ VHH
Recombinant, monoclonal bivalent anti-Vimentin single domain antibody (sdAb) fragment

Which Nano-Booster and Nano-Label conjugates are recommended for super-resolution microscopy?

Nano-Boosters and Nano-Labels are highly suitable for Super-Resolution Microscopy. Due to their small size (2-3 nm), they minimize the linkage error and provide a more precise and dense staining, than conventional antibodies (15 nm linear dimension. The selection of a Nano-Booster and Nano-Label conjugate depends on your microscope setup and lasers. We recommend for:
- STED: ATTO647N, Abberior STAR 635P
- STORM: Alexa Fluor 647, ATTO488
- SIM: ATTO488/594

Are Nano-Labels applicable for live-cell imaging?

Nano-Labels are small proteins and therefore don’t penetrate through non-permeabilized cell membranes. If you need to deliver Nano-Labels into live cells, you may want to apply protein transduction methods (e.g. electroporation) or reagents, however from our experience, the most efficient way is to microinject the Nano-Labels.

Can I do a simultaneous co-staining with two or more Nano-Boosters and Nano-Labels?

Yes, you can combine the Nano-Boosters and Nano-Labels. For example, if you typically use the Nano-Boosters in a 1:200 dilution, you should add 1 µL each of gba488 and rba594 to 200 µL of blocking solution for a co-staining.

How many dye molecules are coupled to Nano-Boosters and Nano-Labels?

Each Nano-Booster and Nano-Label molecule carries on average 1-2 fluorophores.
Note: Nano-Boosters labeled with Atto647N carry a maximum of 1 fluorophore per VHH.

Do Nano-Boosters and Nano-Labels penetrate though the cell membranes of live cells?

No. Nano-Boosters and Nano-Labels are small proteins and therefore don’t penetrate through non-permeabilized cell membranes. If you need to deliver Nano-Booster and Nano-Labels into live cells, you may want to apply protein transduction methods (e.g. electroporation) or reagents, however from our experience, the most efficient way is to microinject the Nano-Boosters and Nano-Labels.

Benefits

  • Specific probe for direct immunostaining of vimentin filaments
  • Higher image resolution and higher labeling density
  • Less than 4 nm epitope-label displacement minimizes linkage error
  • Superior accessibility and labeling of epitopes in crowded cellular/organelle environments
  • Improved target binding due to an avidity effect from the bivalent form of vimentin-Label

 

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Only for research applications, not for diagnostic or therapeutic use!