The dye:protein ratio for live cell imaging has not been confirmed. According to literature references, NTA-Atto 488 for live imaging, used for immunofluorescence or staining His-tagged proteins on cells, typically involves dilutions of 1:200 to 1:1000 of the reconstituted dye (1 mg/mL) in buffer.
39625
NTA - Atto 488
BioReagent, suitable for fluorescence, ≥80% (sum of isomers, HPCE)
Synonym(s):
Atto 488 - Nitrilotriacetic acid, complexed to Ni2+ - ion, Nα,Nα-bis(carboxymethyl)-L-lysine, Nickel(II) complex
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About This Item
NACRES:
NA.32
UNSPSC Code:
12171501
Pricing and availability is not currently available.
product line
BioReagent
Quality Segment
assay
≥80% (sum of isomers, HPCE)
form
solid
fluorescence
λex 501 nm; λem 523 nm in PBS with 0.08% sodium azide
suitability
suitable for fluorescence
detection method
fluorometric
storage temp.
−20°C
General description
find more information here
Application
for the fluorescence detection of HIS-tagged proteins
Legal Information
This product is for Research use only. In case of intended commercialization, please contact the IP-holder (ATTO-TEC GmbH, Germany) for licensing.
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This Item | |||
|---|---|---|---|
| suitability suitable for fluorescence | suitability suitable for fluorescence | suitability suitable for fluorescence | suitability suitable for fluorescence |
| fluorescence λex 501 nm; λem 523 nm in PBS with 0.08% sodium azide | fluorescence λex 554 nm; λem 576 nm in PBS with 0.08% sodium azide | fluorescence λex 647 nm; λem 669 nm in PBS | fluorescence - |
| form solid | form lumps | form - | form powder |
| Quality Level 100 | Quality Level 100 | Quality Level 100 | Quality Level 100 |
| assay ≥80% (sum of isomers, HPCE) | assay ≥80% (sum of isomers, HPCE) | assay ≥80% (HPCE) | assay ≥90% (HPLC), ≥90% (degree of coupling) |
| storage temp. −20°C | storage temp. −20°C | storage temp. −20°C | storage temp. −20°C |
signalword
Danger
Hazard Classifications
Aquatic Acute 1 - Aquatic Chronic 1 - Carc. 1A - Muta. 2 - Repr. 1A - Resp. Sens. 1 - Skin Sens. 1 - STOT RE 1 Inhalation
Storage Class
6.1C - Combustible acute toxic Cat.3 / toxic compounds or compounds which causing chronic effects
wgk
WGK 3
flash_point_f
Not applicable
flash_point_c
Not applicable
ppe
Eyeshields, Gloves, type N95 (US)
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What is the dye:protein ratio used for live cell imaging?
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Can this reagent detect Histidine affinity tag (HAT)? What is the dilution ratio for WB or IF?
1 answer-
This product is provided as 250 micrograms of powder per vial. The contents of the vial can be reconstituted in 250 microliters of phosphate buffered saline (PBS) to make a 1 mg/mL stock solution. Dilutions of 1:1000 to 1:2500 in PBS with TWEEN 20 (PBS-T; Product P3563 at 1X) can be used to stain histidine-tagged proteins by Western Blot (WB).
Immunofluorescence (IF) is typically less sensitive than WB, so dilutions of 1:200 to 1:1000 may be appropriate.
See: https://www.sigmaaldrich.com/US/en/technical-documents/technical-article/protein-biology/protein-labeling-and-modification/ni-nta-atto-conjugatesHelpful?
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Is this product suitable for detection of polyhistidine tagged proteins directly in SDS-PAGE or Western blot?
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ATTO 488 NHS-ester can be used for the fluorescence detection of HIS-tagged proteins and polyhistidine tagged proteins directly in SDS-PAGE or Western blot.
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We have a his-tag labeled recombinant protein that we would like to have fluorescently labeled, before we use it to detect/label antibodies. I came across Ni-NTA-Atto Conjugates and I was wondering wether we could use this to label out his-tagprotein?
1 answer-
There should be no difficulty with the use of the product to label a HIS-tagged recombinant protein as long as the HIS-6 tag is in an acceptable position for binding. Typically, this is done with either a N or a C-terminal His6 tag. Please note that the only tests that we conduct on this material include a solubility, purity and a luminescence assay. Hence, we do not offer a detailed protocol for use of the product for HIS-tag protein labeling.
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What is the number of nickel ions present in product 39624? Is it mono-nta, tris-nta, or another form?
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There is one nickel ion per molecule, but the exact structure is proprietary.
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What is the molecular weight of this product NTA-ATTO488?
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The molecular weight of this product is 964 Da.
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