Histone-Label

Description
Histone-Label* is a fluorescent chromatin probe for direct staining of chromosomes and nuclei. It consists of an anti-histone VHH conjugated to the fluorescent dye ATTO488.
For immunofluorescence microscopy of histones, chromosomes, and nuclei.

Conjugate
This primary Nanobody is available as conjugation with ATTO488.

Specificity
Histone H2A-H2B heterodimers

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

Product Size Code Price Buy
Product Histone-Label Atto488 Size 10 µL Code tba488-10 Price $ 100
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Product Histone-Label Atto488 Size 100 µL Code tba488-100 Price $ 365
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Product Histone VHH, recombinant binding protein Size 250 µL Code tt-250 Price $ 275
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Coupled Nanobody/ VHH
Recombinant, monoclonal anti-histone single domain antibody (sdAb) fragment

Specificity
Histone-Label immunostains chromatin throughout the cell cycle (interphase chromatin, chromatin condensation and mitotic chromosomes) due to specific binding to histone H2A-H2B heterodimers.

Host/isotype
Alpaca/recombinant VHH domain, monoclonal

Available conjugates
ATTO488

Recommended dilution
IF/ICC: 1:400
Optimal working concentration is application-dependent and should be determined by testing a range of dilutions from 1:200 to 1:1,000.

Microscopy techniques
Wide-field or epifluorescence microscopy; confocal microscopy; super-resolution microscopy e.g. 3D-SIM, PALM, STED, STORM; light sheet microscopy

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

Size
10 μL, 100 μL

Protein concentration
0.5 – 1 mg/mL (conjugates)

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

Validation
Tested in cell culture human HeLa cells.
Fixed (formaldehyde, methanol, or glutaraldehyde) and permeabilized cells

License
*Commercialized under license of INRA, the French National Institute for Agricultural Research

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 histones, chromosomes, and nuclei
  • Labels histones throughout the nuclei
  • Better penetration into densely packed nuclei
  • Higher image resolution and higher labelling density
  • Monovalent VHHs do not cluster their epitopes

 

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