Dnmt1-Chromobody plasmids

Description
DNA plasmid encoding for anti-Dnmt1 VHH (anti-Dnmt1 Nanobody) fused to TagRFP or TagGFP2

Specificity
Dnmt1 (DNA (cytosine-5)-methyltransferase 1), tested in human cells

Applications

Live cell imaging
Observing endogenous Dnmt1 localization
Dnmt1 as biomarker during cell cycle progression

Dnmt1-Chromobody plasmid
Dnmt1-Chromobody plasmid (TagGFP2)
Dnmt1-Chromobody plasmid (TagRFP)

Product Size Code Price Buy
Product Dnmt1-Chromobody® plasmid (TagGFP2) Size 20 µg Code dcg Price $ 999
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Product Dnmt1-Chromobody® plasmid (TagRFP) Size 20 µg Code dcr Price $ 999
Buy +

Dnmt1-Chromobody formats

 

Dnmt1-Chromobody
plasmid (TagGFP2)

Dnmt1-Chromobody
plasmid (TagRFP)

Vector type

mammalian
expression vector

mammalian
expression vector

Promoter

constitutive CMV IE

constitutive CMV IE

Reporter

TagGFP2

TagRFP

Codon usage

mammalian

mammalian

Selection

Kan/Neo

Kan/Neo

Nuclear localization
signal (NLS)

No

No

Please contact us for more details.

Specificity
DNA (cytosine-5)-methyltransferase 1 (Dnmt1), tested in human cells

Encoded Nanobody/ VHH
Monoclonal anti-Dnmt1 single domain antibody (sdAb) fragment

Sequence
With the Dnmt1-Chromobody plasmid you receive the sequence information of the Alpaca antibody to Dnmt1 fused to TagGFP2 or TagRFP, as well as the full vector sequence.

Microscopy techniques
Wide-field epifluorescence microscopy; confocal microscopy

Transfection
Transfection of Chromobody plasmids into mammalian cells can be done with standard DNA-transfection methods, e.g. lipofection (Lipofectamine 2000® from Thermo Scientific), according to the manufacturer’s protocol for the transfection reagent. Please choose the transfection method that works the best for your cell type.

Storage instructions
Shipped at ambient temperature. Store at -20 °C.

Are Chromobodies constitutively expressed?

Yes, Chromobodies expression is regulated by immediate early promotor CMV. This promotor allows constitutive Chromobody expression.

Do Chromobodies only work in live cells?

Yes, the Chromobody plasmid is only expressed in live cells. Cells should be transfected with the Chromobody plasmid at least over night to observe the Chromobody location signal. Alternatively, cells can be fixed prior to imaging.
Note: We don't recommend fixation of cells for the Histone-Chromobody.

When should I image my cells after transfection with the Chromobody plasmid?

The Chromobody signal is maintained up to 3 days in the cell. However, this also depends strongly on the cell type.
We recommend to image the cells 16-24 hours after transfection.

Do the Chromobodies diffuse through the cell membrane into growth medium?

No, Chromobodies are small proteins being expressed in the cytosol. They are not secreted into the medium and remain in the cell as long as the cell maintains its plasma membrane integrity.

Are Chromobodies fluorogenic or do they only emit fluorescence when bound to a target?

Chromobodies are chimeric proteins consisting of a VHH fused to a fluorescent protein. They maintain their fluorescence regardless of whether they are bound to a target or not.

Can I amplify the Chromobody plasmid in bacteria?

Yes, the Chromobody plasmids can be propagated in E.coli by standard techniques.

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