Organelle Protein Purity Verification Service

The separation and purification of organelles can identify compartment specific proteins. However, the accuracy of this identification depends on the purity of the target organelle and is also influenced by the degree of protein enrichment and contamination. Creative Proteomics can use a variety of methods, such as Western blotting and enzyme assays, to provide you with organelle protein purity verification services, and to ensure your subsequent research on subcellular organelles. If you choose our organelles isolation and proteins purification service, then we will provide purity verification services for isolated organelles or organelle proteins for free.

Advantages of Organelle Protein Purity Verification Service:

• A sufficient level of expertise.
• Multiple purity assessment methods are used in combination to ensure the accuracy of results.
• Customized purity verification methods for your samples and needs.
• Not limited to organelle protein analysis, other protein purity analysis available.

Methods of Protein Purity Verification:

1. Western Bolt

Creative Proteomics has a variety of specific antibodies against subcellular marker proteins, such as histone 3, calnexin, and mitochondrial ascorbate peroxidase. It can be used to determine the purity of various organelles such as the nucleus, mitochondria and endoplasmic reticulum.

2. Chromatography

• Gel Filtration Chromatography

We will carry out two tests: the first is the non-specific measurement of protein (such as 280nm, 220mn absorbance; The second is specific test (enzyme activity test, immunological test, mass spectrometry, multi-angle light scattering test, etc.) to confirm the identity of the analyzed protein.

• Reversed-Phase High-Performance Liquid (RP-HPLC)

This method is simple and fast, and it can flexibly apply many different characteristics to separate affinity proteins. Creative Proteomics Peptides are usually detected based on UV absorbance at 215 ~ 220nm. Gradient elution can be completed within 1h.

3. Mass Spectrometry

Mass spectrometry works by separating proteins or peptides by mass and charge, accelerating them to the detector and creating a unique spectrum for each protein (or protein fragment). Creative Proteomics has Q Exactive Hybrid Quadrupole-Orbitrap Mass Spectrometer, which can provide you with a variety of protein purity analysis services.

4. Enzyme Assays

• The purity of the plasma membrane: compare H⁺ - ATPase activity with the total ATPase activity.
• The purity of the nuclear fraction: detect histone acetyltransferase activity.
• The purity of mitochondria: estimate a number of enzymes, including fumarase, aconitase and cytochrome c oxidase.
• The purity of chloroplasts: assess phosphoribulokinase activity and NADH-dependent glyceraldehyde 3-phosphate dehydrogenase activity.
• The purity of endoplasmic reticulum: measure NADH: cytochrome c reductase activity.

For more enzymatic analysis, please contact us.

We not only provide purity analysis services for organelle proteins, but also provide a variety of protein purity analysis. If you are interested in our protein purity analysis service or want to learn more about it, please contact us.

References

1. Komatsu S, Hashiguchi A. Subcellular Proteomics: Application to Elucidation of Flooding-Response Mechanisms in Soybean. Proteomes, 2018, 6(1): 13.
2. Kappler L, Li J, Häring H U, et al. Purity matters: A workflow for the valid high-resolution lipid profiling of mitochondria from cell culture samples. Scientific reports, 2016, 6: 21107.

* For Research Use Only. Not for use in diagnostic procedures.