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Nickel Colorimetric Assay Kit

Catalog #: K510

In stock

$325.00

Product Details

Cat # +Size K510-100
Size 100 assays
Detection Method Absorbance (300 to 600 nm)
Species Reactivity Mammalian
Applications The assay is a simple method of quantifying Ni²ᶧ in a variety of samples, which gives a linear range of 2 to 50 nmol Nickel containing less than 25 nmol Cobalt.
Features & Benefits • Simple procedure; takes less than ~40 minutes
• Fast and convenient
Kit Components • Nickel Assay Buffer
• Nickel Reagent
• Nickel Chloride Standard (1.0 µmol)
Storage Conditions -20°C
Shipping Conditions Gel Pack
USAGE For Research Use Only! Not For Use in Humans.

Details

Nickel is one of four ferromagnetic elements (symbol Ni, at. Num. 28). Several enzymes depend on nickel for activity, including some ureases, carbon monoxide dehydrogenases (methane forming enzymes which reduce CO₂ to CH₄) and some hydrogenases which allow the production or removal of H2. Most of these activities are found in the archaebacteria. Nickel forms complexes with sulfhydryl compounds with significant absorbance in the UV/visible region in the presence of other ions. BioVision’s Nickel Assay kit provides a simple method of quantifying Ni²ᶧ in a variety of samples. The assay takes advantage of reaction of Ni²ᶧ with mercaptoethanol in borate buffer to form a complex with strong absorbance bands from ~300 to 600 nm. Fe²ᶧ and Co²ᶧ interfere with the assay, therefore extra steps (as described below) must be taken to subtract the interference in order to determine the correct Nickel concentration in mixed samples. Other ions tested (Mn²ᶧ, Cu²ᶧ, Zn²ᶧ) do not interfere with the assay, presumably no other ionic species are present in high enough concentration to interfere with the reaction. The assay is a simple method of quantifying Ni²ᶧ in a variety of samples, which gives a linear range of 2 to 50 nmol Nickel containing less than 25 nmol Cobalt.


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Can this kit be used with samples like bacteria, plants, drosophila, yeast etc?
We have optimized the kit with mammalian samples. However, theoretically these kits should work with samples from multiple species/sources. Since the optimal conditions depend on the sample type, the protocol has to be be adapted to fit the samples for efficient results. Please refer to this kit's citations to see what kind of samples have been used with this kit other than mammalian samples.
Can we use frozen samples with this assay?
Fresh samples are always preferred over frozen samples. However, frozen samples can also be used, provided, they were frozen right after isolation, were not freeze thawed multiple time (for which we recommend aliquoting the samples before freezing) and have been frozen for relatively short periods.
Can we use a different wavelength than recommended for the final analysis?
It is always recommended to use the exact recommended wavelength for the most efficient results. However, most plate readers have flexibility in their band width of detection in increments of +/- 10 nm. Depending on this flexibility range, you can deviate from the recommended wavelengths within limits.
What is the exact volume of sample required for this assay?
There is no specific volume we can recommend for the amount any sample to be used since it is completely sample concentration and quality based. You have to do a pilot expt with multiple sample volumes to determine the optimal volume which gives a reading within the linear range of the standard curve. Please refer to the citations for this product to see what other clients have used with similar sample types.
Do you have trial sizes of this kit?
Unfortunately, we do not have trial sizes of this kit available. However, if you are based in the US or Canada, we will give you a 10% off list price introductory discount on its purchase price. If you are based out of this area please contact yopur regional BioVision distributor.
What is the shelf life of this kit?
This kit is good for 12 months from the date of shipment in the unopened form when stored at the appropriate temperature and appropriate conditions. After opening and reconstitution, some of the components in this kit are good for 2 months at -20°C. Please refer to the datasheet for storage information and shelf life of each of the components.
Why are my standard curve values lower than those shown on the datasheet?
There are multiple factors which influence the signals like the incubation times, room temperature, handling etc. In general, to increase the value of the standards, you can increase the incubation time. As long as the standard curve is linear, it should be fine to use, since all of your samples will also be measured under the same conditions on this curve.
How do I normalize my samples against protein concentration?
You can use a protein quantitation assay on the supernatants you get from cell/tissue lysates or with any other liquid sample in the assay buffer.
Can we purchase individual components of this kit?
Yes, you can purchase any of the kit's components without the whole kit. Please refer to the component Cat #s mentioned on the datasheet for ordering.
Can we use an alternate buffer for sample preparation (cell lysis, sample dilutions etc)?
Our assay buffers are optimized for the reactions they are designed for. They not only contain some detergents for efficient lysis of your cells/tissue, but also contain some proprietary components required for the further reactions. Therefore, we highly recommend using the buffers provided in the kit for the best results.
Should I make a standard curve for every expt I do, or is one curve/kit enough?
Yes, I would strongly recommend you to do the standards every time you do the expt. There is always a chance that something was done differently that day and we do not want any conditions to differ between standards and samples.
Can EDTA, DTT or Imidazole interfere with the assay?
With low concentrations of EDTA it might be fine to use the kit (=1mM). However, with high concentrations of EDTA (concentrations used to elute the protein from Ni-NTA column) can interfere with the assay as it will chelate the metal and there is not enough salt in the system to titrate it down. DTT is not compatible. Very low concentration of Imidazole may not interfere with the assay, but a high concentration like the ones used for washing proteins may interfere with the assay.
Can Fe+3 ions interfere with the assay?
The assay buffer has a reducing agent that can reduce Ferric to ferrous which can affect the assay signal.
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