Human Aspartate Aminotransferase (AST) Protein

Proteins and Peptides

Aspartate Aminotransferase (AST)

293F cells

Activity: Active
Biological Activity: Specific activity: 4.9 U/mg In this assay, an amino group is exchanged from aspartate to alpha-ketoglutarate. The products of this reversible transamination reaction are oxaloacetate and glutamate. The oxaloacetic acid can be decomposed into pyruvate and carbon dioxide in the presence of phenylamine citrate. The activity of aspartate transaminase can be measured by calculating the concentration of the pyruvate. Blank, sample and standard tubes were prepared and labelled. 10 µl of different concentrations of recombinant AST were added to respective sample tubes. 10 µl of phosphate buffer was added to the blank tube in place of AST. The standard curve was prepared by 2-fold dilution of 2 µM pyruvate with phosphate buffer. 50 µl of substrate containing 2 mM 2-Ketoglutaric acid, 0.1 M L-aspartic acid, 0.2 M phosphate buffer, pH 7.4, was added to all vials. All vials were incubated at 37 °C for 1 hour. 10 µl phenylamine citrate and 50 µl 2,4-dinitrophenylhydrazine were added to all vials. The vials were incubated at 37 °C for 20 min. 500 µl of 0.4 M NaOH was added to the blank and sample tubes to stop the reaction, and then the OD value was recorded at 520 nm. One unit of AST Activity (U) is the amount of enzyme that generates 1 μmol of pyruvate per minute (pH 7.4, 37 °C). AST Activity (U/mg) =  OD × d / t × P where:   OD - Absorbance at 520 nm   d - Dilution factor   t - Reaction time (min)   P - Quantity of protein
Sequence Fragment: Met1-Gln413

Human

Unconjugated

Calculated MW: 47.8 kDa  Observed MW (SDS-PAGE): 44 kDa

Recombinant

> 97%

50 µg

100 µg

200 µg

500 µg

1 mg

Lyophilized

WB, SDS-PAGE

Prior to lyophilization: PBS, pH 7.4, containing 0.01% Sarcosyl, 1 mM DTT, 5% Trehalose and Proclin-300.

Shipped within 5-7 working days.

Store lyophilized form at 2-8°C for up to 1 month. For longer periods, store lyophilized or liquid at -80°C. Avoid repeated freeze–thaw cycles.

No

P17174

6609

607608

Protein AST

RUO

THIS PRODUCT IS FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC, THERAPEUTIC OR COSMETIC PROCEDURES. NOT FOR HUMAN OR ANIMAL CONSUMPTION.
To keep the original salt concentration, we recommend reconstituting to the original concentration prior to lyophilization (see Concentration) in ddH2O. If a lower concentration is required, dilute in 10 mM PBS, pH 7.4. If a higher concentration is required, the product can be reconstituted directly in 10 mM PBS, pH 7.4, though please note that this will change the overall salt concentration. The stock concentration should be between 0.1-1.0 mg/ml. Do not vortex.
Concentration: Prior to lyophilization: 200 µg/ml