Asparagine Assay Kit (Fluorometric) (ab273333)
Key features and details
- Assay type: Quantitative
- Detection method: Fluorescent
- Platform: Microplate
- Sample type: Adherent cells, Cell culture media, Other biological fluids, Suspension cells
Overview
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Product name
Asparagine Assay Kit (Fluorometric) -
Detection method
Fluorescent -
Sample type
Other biological fluids, Adherent cells, Suspension cells, Cell culture media -
Assay type
Quantitative -
Assay duration
Multiple steps standard assay -
Product overview
Asparagine Assay Kit (Fluorometric) (ab273333) allows for highly sensitive quantification of L-asparagine levels in biological fluids and cultured cells.
In the assay, L-asparagine is hydrolyzed to generate aspartate, which is converted to pyruvate and subsequently reacts with a fluorogenic probe to form a stable fluorophore (Ex/Em = 538/587 nm) via a series of coupled enzymatic reactions. The assay is not affected by physiological concentrations of other common amino acids and can detect asparagine levels down to 5 μM in samples. -
Platform
Microplate
Properties
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Storage instructions
Store at -20°C. Please refer to protocols. -
Components 100 tests Asparagine Assay Buffer 1 x 25ml Asparagine Enzyme Mix Lyophilized 1 vial Conversion Enzyme Mix Lyophilized 1 vial Developer Enzyme Mix Lyophilized 1 vial L-Asparagine Standard Lyophilized 1 vial Probe Solution 1 x 200µl Sample Cleanup Mix Lyophilized 1 vial
Images
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Example data
Example data.
L-Asparagine Standard curve.
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Example dataExample data.
Specificity for detection of L-asparagine (ASN) over other common L-amino acids. At a 5- fold molar excess (10 nmole/well) versus ASN (2 nmole/well), tryptophan (TRP) gives ~5% interference and the reduced form of cysteine (CYS) gives ~20% interference (the oxidized disulfide form, which accounts for 90% of the total cysteine in biological fluids, generates ≤1% interference at 5-fold excess).
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Example dataExample data.
Estimation of total asparagine in pooled heparinized human plasma (10μl), pooled off-the-clot human serum (10μl) and pooled human CSF (15μl). Asparagine concentrations for plasma, serum and CSF samples were 25.61 ± 2.51 μM, 37.99 ± 1.77 μM and 11.11 ± 2.59 μM, respectively. Data are mean ± SEM of at least 3 replicates, samples were treated with Sample Cleanup Mix, deproteinized using 10 kDa spin columns and assayed according to the kit protocol.