Recombinant Human CES1 (carrier-free)

Product Details

Source

Human CES1, amino acid (His19 – Glu563) (Accession # AB119996.1) was expressed in CHO cells. The C-terminus contains a 6His-GGQ tag.

Molecular Mass

The 558 amino acid recombinant protein has a predicted molecular mass of approximately 61.5 kD. The DTT-reduced and non reduced protein migrates at approximately 70 kD and 60 - 80 kD respectively by SDS-PAGE. The C-terminus contains a 6His-GGQ tag. The predicted N-terminal amino acid is His.

Purity

> 95%, as determined by Coomassie stained SDS-PAGE

Formulation

0.22 µm filtered protein solution is in 50 mM Sodium Acetate, 150 mM NaCl, pH 5.0.

Endotoxin Level

Less than 0.1 EU per µg cytokine as determined by the LAL method

Concentration

25 µg size is bottled at 200 µg/mL. 100 µg size and larger sizes are lot-specific and bottled at the concentration indicated on the vial. To obtain lot-specific concentration and expiration, please enter the lot number in our Certificate of Analysis online tool.

Storage & Handling

Unopened vial can be stored at -20°C or -70°C for six months. For maximum results, quick spin vial prior to opening. Avoid repeated freeze/thaw cycles.

Activity

Human carboxylesterase 1 (CES1) hydrolyzes the compound, 4-nitrophenyl acetate in a colorimetric assay. The specific activity is > 6,000 pmol/min/µg in the presence of 0.04 µg of recombinant human CES1.

Application

Bioassay

Application Notes

Human CES1 Enzymatic Assay

Human carboxylesterase 1 hydrolyzes the carboxylic acid substrate, 4-nitrophenyl acetate. The increase of hydrolyzed product is monitored by an increase in absorbance at 400 nm. The specific activity is > 6,000 pmol/min/µg in the presence of 0.04 µg of recombinant human CES1.

Materials

• Assay buffer: 50 mM Tris, pH 7.5
• Recombinant human CES1
• Substrate: 4-nitrophenyl acetate (4-NPA) (Sigma, Cat. No. N-8130), 100 mM stock in acetone
• 96-well clear plate (Costar, Cat. No. 92592)
• Plate reader (Model: SpectraMax Plus by Molecular Devices or equivalent)

​Procedure

1. Dilute the recombinant human CES1 to 0.8 µg/mL in assay buffer.
2. Dilute the substrate to 2 mM in deionized water.
3. In a plate load 50 μL of human CES1 to the wells and start the reaction by adding 50 µL of 2 mM substrate. Include a substrate blank containing 50 µL of substrate and 50 µL assay buffer.
4. Read at a wavelength of 400 nm (bottom read) in kinetic mode for 5 minutes.
5. Calculate specific activity: 

     Specific Activity (pmol/min/µg) = Adjusted Vmax* (OD/min) x Conversion Factor** (pmol/OD)                                                                                             amount of enzyme (µg)

*Adjusted for substrate blank
**Derived using calibration standard 4-Nitrophenol (Sigma, Cat. No. 241326)
Per Well:
Recombinant human CES1: 0.04 µg
Substrate: 1 mM

BioLegend carrier-free recombinant proteins provided in liquid format are shipped on blue ice. Our comparison testing data indicates that when handled and stored as recommended, the liquid format has equal or better stability and shelf-life compared to commercially available lyophilized proteins after reconstitution. Our liquid proteins are validated in-house to maintain activity after shipping on blue ice and are backed by our 100% satisfaction guarantee. If you have any concerns, contact us at tech@biolegend.com.

Antigen Details

Structure

Homotrimer

Distribution

Liver, human lung, and in monocytes/macrophages, absent in intestine

Function

Drug clearance, activation, and deactivation. Hydrolysis of exogenous and endogenous compounds such as carboxylic acid esters, thioesters, carbamates, and amides.

Interaction

Xenobiotics and fatty acids, cholesterol

Bioactivity

Human carboxylesterase 1 (CES1) hydrolyzes the carboxylic acid substrate, 4-nitrophenyl acetate. The increase of hydrolyzed product is monitored by an increase in absorbance at 400 nm.

Antigen References

1. Burgaleta C, et al. 1999. Cancer. 86:2133.
2. Oertel J, et al. 1985. B J Haem. 61:717.
3. Redinbo MR, et al. 2003. Biochem Soc Trans. 31:620.
4. Satoh T, Hosokawa M, 2006. Chem Biol Interact. 162:195.
5. Wang X, et al. 2017. Pharmacogenomics J. 16:2220
6. Orcholski ME, et al. 2017. Am J Physiol Lung Cell Mol Physiol. 313:L252.
7. Wang X, et al. 2019. Proteomics. 19:e1800288.

Gene ID

1066 View all products for this Gene ID

UniProt

View information about CES1 on UniProt.org

Documentation

• Technical Data Sheet (pdf)
• Certificate of Analysis
• Material Safety Data Sheet

Reviews

Related Pages & Pathways

Related FAQs

Why choose BioLegend recombinant proteins?

     • Each lot of product is quality-tested for bioactivity as indicated on the data sheet.
     • Greater than 95% Purity or higher, tested on every lot of product.
     • 100% Satisfaction Guarantee for quality performance, stability, and consistency.
     • Ready-to-use liquid format saves time and reduces challenges associated with reconstitution.
     • Bulk and customization available. Contact us.
     • Learn more about our Recombinant Proteins.

How does the activity of your recombinant proteins compare to competitors?

We quality control each and every lot of recombinant protein. Not only do we check its bioactivity, but we also compare it against other commercially available recombinant proteins. We make sure each recombinant protein’s activity is at least as good as or better than the competition’s. In order to provide you with the best possible product, we ensure that our testing process is rigorous and thorough. If you’re curious and eager to make the switch to BioLegend recombinants, contact your sales representative today!

What is the specific activity or ED50 of my recombinant protein?

The specific activity range of the protein is indicated on the product datasheets. Because the exact activity values on a per unit basis can largely fluctuate depending on a number of factors, including the nature of the assay, cell density, age of cells/passage number, culture media used, and end user technique, the specific activity is best defined as a range and we guarantee the specific activity of all our lots will be within the range indicated on the datasheet. Please note this only applies to recombinants labeled for use in bioassays. ELISA standard recombinant proteins are not recommended for bioassay usage as they are not tested for these applications.

Have your recombinants been tested for stability?

Our testing shows that the recombinant proteins are able to withstand room temperature for a week without losing activity. In addition the recombinant proteins were also found to withstand four cycles of freeze and thaw without losing activity.

Does specific activity of a recombinant protein vary between lots?

Specific activity will vary for each lot and for the type of experiment that is done to validate it, but all passed lots will have activity within the established ED50 range for the product and we guarantee that our products will have lot-to-lot consistency. Please conduct an experiment-specific validation to find the optimal ED50 for your system.

How do you convert activity as an ED50 in ng/ml to a specific activity in Units/mg?

Use formula Specific activity (Units/mg) = 10^6/ ED50 (ng/mL)

Certificate of Analysis