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InformationUBIQUITIN (Ub) 10-4000pg/ml Assay Range ELISA KIT (Part hUb-ELISA, HUMAN)
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Ubiquitin ELISA Kits (96T)
Ubiquitin ELISA Kits (96T)
Dynamic Assay Range: 10pg/ml - 4,000pg/ml depending on species.
Sensitivity: 5.13 pg/ml.
Lead Time: Approximately 1.5 weeks
Fully guaranteed to meet specifications
Ubiquitin ELISA Kit Specifications
| Part |
Species |
Standard |
Assay Range |
Price (96T) |
| hUB-ELISA | Human | 4800pg/ml | 10pg/ml - 4000pg/ml | $635 |
| rUB-ELISA | Rat | 4800ng/L | 10ng/L - 4000ng/L | $635 |
| mUB-ELISA | Mouse | 3200ng/L | 10ng/L - 3000ng/L | $635 |
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Highlights
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Introduction
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Ubiquitin is a post translational modification to biomolecules targeted to degradation: Attachment of polyubiquitin to a protein targets the protein for degradation in the ubiquitin-proteasome pathway. The addition of ubiquitin can also code for changes in cellular locatization, activity or modulate protein interactions. Ubiquitination is controlled in three main steps; activation, conjugation and ligation, which are performed by ubiquitin-activating enzymes (E1s), ubiquitin-conjugating enzymes (E2s) and ubiquitin ligases (E3s), respectively. The result of this sequential cascade is the conjugation of ubiquitin to lysine residues via an isopeptide bond or to the amino group of the protein's N-terminus via a peptide bond. Ubiquitinated protiens can be transmembrane, cytosolic or nuclear.
Isolating ubiquin-conjugated proteins for ELISA Assay
Ubiquitinated proteins are located in the cell membrane, in the cytosol and in the nucleus. Cell lysis in a non-denaturing buffer is the general procedure employed to access proteins in these cellular compartments. The Fivephoton Biochemicals ELISA Protein Lysis Buffer (Part ELSP-1) is applicable toward isolating a total cellular pool of ubiquitinated proteins for ELISA, while the Fivephoton Biochemicals Nuclear-Cytoplasmic Fractionation Kit (Part NPI-1) can be employed to fractionate cytosolic and nuclear pools of CSF, serum and plasma.
Methods Overview
The included ELISA micro-strip plate is provided pre-coated with an a monoclonal capture antibody selective to the antigen. Samples are pre-mixed with a biotin-linked detection antibody and streptavidin-HRP, which are then added to the plate for 60 min. The plate is washed after the 60 min incubation period. Color reagents are added, resulting in a reaction with HRP that yields a blue colored solution. A stop solution is applied to terminate the reaction, and the solution is measured at 450n
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Sandwich ELISA Methods to Ubiquitin
ELISA Assay Steps
- Mix sample, biotinylated-detection antibody and streptavidin-HRP.
- Apply sample and antibody-HRP complex to ELISA plate for 60min at 37oC. Wash 5 times.
- Apply chromogens, incubate, then stop reaction.
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Product Citation
Goetzl, Edward J et. al. 2015. Altered lysosomal proteins in neural-derived plasma exosomes in preclinical Alzheimer disease.
Neurology July 7, 2015 vol. 85 no. 1 40-47
General Refernces on Ubiquitin
1. Hershko, Avram, and Aaron Ciechanover. 1998. "The ubiquitin system." Annual review of biochemistry 67
2. Leroy, Elisabeth, et al. 1998. "The ubiquitin pathway in Parkinson's disease." Nature 395.6701, 4
3. Mori, Hiroshi, Jun Kondo, and Y. A. S. U. O. Ihara. 1987. "Ubiquitin is a component of paired helical filaments in Alzheimer's disease." Science 235.4796, 1641-1644.
For research use only, not for therapeutics or diagnostics.
Safety: Stop solution contains acid. Avoid ingestion, skin and eye contact.
Storage: 4oC, 6 months.
Shipping: Overnight shipping recommended. Also available for international delivery.
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